DE102004038076A1 - Use of annexin A3, or other proteins, as diagnostic markers for cancer, especially of the prostate, also use of modulators of these markers for treating cancer - Google Patents

Abstract

Use of the protein annexin A3 (I) as a diagnostic marker for prostatic cancer (PC), is new. Independent claims are also included for the following: (1) use of mitochondrial enoyl-coenzyme A-hydratase (III), ubiquitin-isopeptidase T (IV) and/or protein disulfide isomerase (V), serum amyloid P-component (VI), HES1 (VIII), the proteosome alpha -2 subunit (IX) or both (IV) and VI) as diagnostic markers for cancer; (2) use of nuclear chloride ion channel protein (VIII), adenine-phosphoribosyl transferase (X) and inorganic pyrophosphatase (XI) as diagnostic markers for PC; (3) use of a combination of the specified proteins as markers for cancer; (4) diagnostic kit that includes at least one substance for determining activity and/or abundance of the specified proteins; (5) use of an agent (II) that interacts with one of the specified marker proteins to modify its activity and/or abundance, to prepare a composition for treating (prostatic) cancer; and (6) screning for (II) that uses at least one of (I), (IV), (VI), (VII), 14-3-3 protein tau, (III), (VIII), (IX), (X), (XI) and/or their dervatives. ACTIVITY : Cytostatic; Osteopathic; Antiarthritic; Antiarteriosclerotic. No biological data is given. MECHANISM OF ACTION : Regulating activity and/or expression of marker proteins.

Description

The The present invention relates to the use of various proteins as diagnostic markers for cancer, the use of drugs for the treatment of cancer and related pharmaceutical Compositions and kits.

cancers are generally due to the emergence of one or more Tumors labeled. Under tumor is a tumor or locally circumscribed Increased tissue volume understood. In a broader sense, each one can localized swelling, z. By edema, acute and chronic Inflammation, Aneurysmal enlargement, an inflammatory organ swelling (z. B. as a so-called spleen tumor). In the narrow sense become under tumor tissue neoplasms (plant, plastome, neoplasia) in Form of a spontaneous, at different degrees uninhibited, autonomous and irreversible excess growth from the body's own Tissues, usually with different from embossed loss specific cell and tissue function is understood.

• I. nach ihrem biologischen Verhalten: 1. benigne (gutartige) Tumore mit differenzierten Zellen und langsamen, lokal verdrängendem Wachstum. 2. maligne (bösartige) Tumore mit Zellkernpolymorphie, Zellatypie, Anaplasie und infiltrierendem, meist raschem, destruierendem Wachstum und Metastasierung. 3. semimaligne Tumore mit den histologischen Kennzeichen maligner Tumore und lokal infiltrierendem Wachstum, jedoch in der Regel ohne eine Metastasierung.
• II. histogenetische Systematik: Hierbei werden die Tumore klassifiziert anhand des Gewebes, aus dem sie entwicklungsgeschichtlich hervorgegangen sind. Es gibt: 1. epitheliale Tumore, die aus Ektoderm und Entoderm hervorgegangen sind: a) benigne Tumore wie z. B. Adenom, Papillom und Polypen. b) maligne Tumore wie z. B. Karzinom. 2. mesenchymale Tumore, hervorgegangen aus dem Mesoderm: a) benigne Tumore wie z. B. Lipom, Fibrom, Osteom, Myom, Leiomyom, Rhabdomyom, Chondrom, b) maligne Tumore wie z. B. die Sarkome. 3. embryonale Tumore sind aus undifferenziertem Gewebe hervorgegangen. Hierzu zählen z. B. Nephroblastome, Neuroblastome, Medulloblastome, Retinoblastome sowie embryonale Rhabdomyosarkome und Teratome.
• III. Klassifikation nach klinischen und pathologischen Befunden: Unter anderem gelten hier die TNM-Klassifikation, Grading, Laurén-Klassifikation, Dukes-Klassifikation, Kieler-Klassifikation, Rappaport-Klassifikation etc.

The tumors are subdivided for better classification into:

• I. Biological behavior: 1. Benign (benign) tumors with differentiated cells and slow, locally displacing growth. 2. Malignant tumors with nuclear polymorphism, cell atypia, anaplasia and infiltrating, mostly rapid, destructive growth and metastasis. 3. Semimalignant tumors with the histological features of malignant tumors and locally infiltrating growth, but usually without metastasis.
• II. Histogenetic systematics: The tumors are classified on the basis of the tissue from which they originated in the history of development. There are: 1. Epithelial tumors, which have emerged from ectoderm and endoderm: a) benign tumors such. Adenoma, papilloma and polyps. b) malignant tumors such. B. Carcinoma. 2. mesenchymal tumors arising from the mesoderm: a) benign tumors such. As lipoma, fibroma, osteoma, myoma, leiomyoma, rhabdomyoma, chondroma, b) malignant tumors such. As the sarcoma. 3. embryonic tumors have emerged from undifferentiated tissue. These include z. Nephroblastomas, neuroblastomas, medulloblastomas, retinoblastomas and embryonic rhabdomyosarcomas and teratomas.
• III. Classification according to clinical and pathological findings: Among others, the TNM classification, grading, Laurén classification, Dukes classification, Kiel classification, Rappaport classification, etc. apply here.

Nice this short overview the tumor division shows what diversity (and partly conflict) exists within the various types of tumors. So z. Eg not only to distinguish between benign and malignant tumors, but also between mortality or lethality of the individual tumors and the likelihood of getting one benign tumor evolved into a malignant tumor.

Separate Tumors such. Mammary carcinomas (breast cancer), the most common malignant tumor of the woman, occur mostly between the 45th and 70th year on. early symptoms are suspicious Palpation findings, usually as a result of cancer screening tests as well as with regular self-examination the breast are discovered. Dependent of tumor stage and degree of differentiation of the tumor can thereby the prognosis from quite positive to very bad. As a result the early one lymphogenous and hematogenous Metastasis in breast cancer is a matter of rapid diagnosis of the tumor, as early as possible with to use the therapy.

Prostate carcinoma (carcinoma of the prostate), in contrast, is the most common malignant tumor of the man, which occurs mainly between the 50th and 70th year of life. The majority are adenocarcinomas. This malignant tumor spreads first through infiltrating growth within the prostate, later an infiltration of seminal vesicles and pelvic connective tissue occurs, relatively rarely also of the rectum, urinary bladder or urethra. Metastasis is lymphogenous and / or hematogenous. Depending on the degree of histological differentiation and the clinical stage, therapy is generally performed by radical prostatectomy with regional lymphadenectomy, and in the advanced stage withdrawal of male sex hormones. Again, the prognosis depends on the stage of the carcinoma. While in a very early stage after a radical prostatectomy in about 90% of cases, a healing occurs at advanced stage rather with a pessimistic prognosis.

prostate cancers are distinguished from prostatic hyperplasia in the diagnosis. at prostate hyperplasia is a benign, so benign tumor. This increases the prostate by numerical increase in the cells and glands of the Stroma. Prostatic hyperplasia is the most common cause of bladder emptying disorders Men. Clinically it starts mainly between the 40th and 50th year of life. The course is slow and in turns. The appearance of complaints Usually takes place only after years with gradual weakening of the Urinary stream and delayed Miktionsbeginn. This can be used as a therapy or alleviation of the symptoms the administration of phytotherapeutic agents should be considered.

There generally an early one Recognition, d. H. Diagnosis, the tumors important for rapid onset of therapy and the prognosis is better, the earlier the Tumor is a set of so-called tumor markers in clinical use. Tumor markers are generally substances and cellular changes whose qualitative or quantitative analysis is a statement about Vor, course or prognosis of (malicious) Diseases allow can. Tumor markers are divided into:

1. Cellular Tumor Markers:

among them among others, cell-membrane-borne tumor antigens, receptors (eg hormone receptors, receptors for growth-promoting Substances in leukemia) and cell markers indicative of increased expression of oncogenes and a monoclonal cell growth, as well as molecular genetic cellular changes, especially chromosome aberrations.

2. Humoral Tumor Markers:

These are opposite physiological conditions in body samples, especially in serum, urine and other body fluids in elevated concentrations detectable (usually physiologically occurring) substances released by the Tumor tissue synthesized and / or secreted by tumor breakdown released or formed as a reaction of the organism to a tumor become. The physiological significance of tumor markers is insufficient known. In the human organism, they usually do not work immunogenic. The clinical (diagnostic) significance depends on their specificity and sensitivity. The humoral tumor markers are divided into two groups. In In the first group the humoral tumor markers are summarized be produced by the tumor itself. These include z. B. tumor associated Antigens, certain hormones (eg gastrin, cortisol etc.), enzymes (eg, neuron-specific enolase (NSE)), as well as proteins (eg, Bence-Jones protein). In the second group contains the tumor markers that are from the tumor Although induced, but not self-produced. Important humoral Tumor markers of this group are z. B. alkaline phosphatase (AP), LDH, Neopterin etc.

The statements made so far show how important selective and sensitive Detection method for Tumors are. Furthermore, there is a great need for new targets in tumor or cancer therapy.

Accordingly The object of the invention is to provide new diagnostic markers for cancer as well as to provide new targets and agents for cancer therapy.

These Task is through the objects the independent one claims solved. Preferred embodiments are in the dependent claims called. The wording of all claims is hereby incorporated by reference into the content of the description.

It could through intense comparative analysis between malignant degenerate Tissue (cancerous tissue) and non-degenerated tissue different proteins be identified, which in these tissues in their frequency or concentration (abundance) differ significantly. The characteristic Abundance of a given protein compared with controls therefore an important indicator for the presence of degenerate cell growth, ie cancerous tissue, dar. According to the invention These proteins are used as diagnostic markers for cancer.

To identify these proteins, samples from cancer tissue (prostate cancer) and healthy prostate tissue were prepared and each of the two samples labeled with different radioactive isotopes. The samples were mixed together and electrophoresed together on a two-dimensional polyacrylamide gel. The signals of each isotope were separately detected and the corresponding protein spots further analyzed. With this procedure, it was possible to identify various proteins that differ significantly in their frequency in the cancerous tissue from the frequency in the healthy tissue. Some of the different proteins are present in cancerous tissue much more frequently, ie upregulated, and sometimes significantly less present, that is downregulated.

The Invention the use of the protein annexin, in particular annexin A3, as diagnostic marker for Cancer. The inventors were able to show that this protein is specific Patient groups in cancerous tissue is up-regulated 5-fold. Therefore is preferably a downregulation of this protein in comparison examined with controls as a characteristic feature of cancerous tissue. The Annexins are a family of structurally related proteins, the phospholipids depending can bind calcium and form calcium pores. The exact role of Annexine is so far unclear.

It is believed that the Annexins both in intracellular as well as extracellular Involved in operations are. However, it is unknown how annexin secretes become. For example, they do not have classical leader sequences for one Translation into the lumen of the endoplasmic reticulum. Da annexines but partly in small secreted vesicles, the so-called Exosomes were found to be annexins by lysis this exosomes the exterior of the Cell can reach. Generally, exosomes play a role in antigen presentation in the immune system Role and are involved in the MHC class I / T cell system.

Interestingly, annexins are involved in the process of bone mineralization (Wang, W., Xu, J., Kirsch, T. 2003, Annexin-mediated Ca ²⁺ influx regulates growth plate chondrocyte maturation and apoptosis., J. Biol. Chem. 278: 3762-9). This is particularly noteworthy because prostate cancer metastases cause an unusually high frequency of osteoblastic bone lesions compared to other cancers. Most cancer metastases are characterized by an osteolytic activity, ie by a bone degradation, prostate cancer metastases, however, show both osteoclastic, so degrading, and osteoblastic, ie bone-building, activity. For this purpose, normal bone crystals are first broken down and then rebuilt as disordered bone deposits. Although this mechanism is poorly understood, physiological mineralization processes play a role here. Mineralization is initiated by small vesicles, called matrix vesicles, which release skeletal cells (osteoblasts) that mineralize from plasma membranes. The first mineral phase (calcium phosphate crystals) forms within the matrix vesicles. Because these vesicles are surrounded by membranes, channel proteins are needed to allow the minerals to penetrate into the vesicles. Important components of these vesicles are the proteins annexin II, V and VI as well as collagen type II and X on the outer surface of the vesicles, which are attached to the outer vesicle surface via binding to annexin V. The annexins form channels in the membranes of the matrix vesicles, through which Ca ²⁺ penetrates into the vesicles. Collagen bound to annexin V enhances these channel activities and mediates, with other annexins, rapid Ca ²⁺ influx and formation of the first crystalline phase within the vesicles. This leads to the initiation of mineralization. When the intracellular crystals reach a certain size, they destroy the membrane and lyse the vesicles. The crystals continue to grow (growth phase of mineralization) and contribute to bone formation. According to the results of the inventors, this role of annexins in the abnormal bone mineralization by prostate cancer metastases is presumably linked to the upregulation of annexin A3 in cancerous tissue. In this context, inorganic pyrophosphatase which releases inorganic phosphate and which is up-regulated according to the inventors' results in cancer, in particular in prostate cancer, can also be seen. From the upregulation of annexin A3 in cancer cells, it can be concluded that annexin A3 plays a biological role in prostate cancer cell exosomes. For this purpose, especially a connection with ion channels comes into question. In a preferred use of the protein annexin A3, therefore, the activity of the protein in exosomes is influenced. This preferably leads to a changed immune surveillance of the tumor cells. Since extracellular annexin A3 has a higher concentration in the vicinity of the tumor cells, an affinity reagent, in particular an antibody which has a high affinity for annexin A3, can be used to deliver drugs such as toxins or radioactive doses to the vicinity of the tumor , Such an agent should preferably not effectively cross the cell membrane, so that advantageously healthy cells which express only intracellular annexin A3 are unaffected thereby. Interestingly, matrix vesicles were also observed in association with osteoarthritic cartilage and arteriosclerotic lesions.

In a particularly preferred use of the protein annexin A3 upregulation of this protein in combination with downregulation Annexin A1, Annexin A2 and / or Annexin A5 examined. This is preferably done in comparison with controls. It already became shown that annexin A1, A2 and annexin A5 in cancerous tissue and especially in prostate cancer tissue is down regulated. Hence, an analysis of the downregulation is of annexin A3 in combination with downregulation of one or more several of these other annexins for a diagnosis particularly revealing.

The Invention continue the use of the protein ubiquitin isopeptidase T and / or the protein Protein disulfide isomerase (PDI) as a diagnostic marker for cancer. This is advantageously a down-regulation of the ubiquitin isopeptidase T and / or upregulation of the protein disulfide isomerase (PDI) in comparison with controls as a characteristic feature for cancerous tissue used. The inventors were able to show that the ubiquitin isopeptidase T about 5 x less abundant in cancerous tissue and the PDI about twice expressed as high in cancer tissue as compared with healthy tissue becomes. This shows an inverse correlation between PDI and Ubiquitin isopeptidase T.

The Ubiquitin isopeptidase T is an enzyme which, among other enzymes at the ubiquitin-dependent Proteolysis of proteins is involved. After addition of a polyubiquitin chain to the target protein, the ubiquitinated protein is transformed by a complex, consisting of many subunits, known as 26 S-proteasome is mined. The subsequent detachment of the polyubiquitin chain is mediated by the zinc-binding enzyme ubiquitin-isopeptidase T. Like the ubiquitin isopeptidase T is also the PDI in the controlled proteolysis of proteins, thus involved in apoptotic processes. PDI interacts in the endoplasmic reticulum under certain conditions with ubiquitin, which has a ubiquitin-like domain and a ubiquitin-associated one domain having. This association has already become functional Related to obtaining a tolerance to ischemic Stress and Apoptosis (Ko H.S. et al., 2002, J. Biol. Chem. 277: 35386-92).

These Relationship to apoptotic processes between both enzymes makes an observation of the downregulation of both proteins as a characteristic feature of Cancer tissue particularly suitable. On the other hand, it is also beneficial if only one of these proteins, especially the ubiquitin isopeptidase T, in which abundance is analyzed as a diagnostic feature. This is particularly vor geous because the ubiquitin isopeptidase T in their frequency in cancerous tissue very clearly compared with controls, d. H. on about a fifth, is reduced.

Farther comprises the invention involves the use of mitochondrial enoyl-coenzyme A hydratase as diagnostic marks for Cancer. This protein can also be used in combination with the fatty acid-binding Protein 3 (FABP-3) and / or the epidermal fatty acid-binding Protein (E-FABP) can be used. It is particularly preferred here, if up-regulation of mitochondrial enoyl-coenzyme A hydratase and / or of the epidermal fatty acid-binding Protein (E-FABP) and / or Downregulation of Fatty Acid-Binding Protein 3 (FABP-3) is examined in comparison with controls, since according to the invention such Upregulation or downregulation of these proteins as characteristic Characteristic for Has exposed cancerous tissue.

The Inventors were able to show that the mitochondrial enoyl-coenzyme A hydratase in cancerous tissue is about 2.8 times increased in its abundance. This enzyme has already been linked to β-oxidation of fatty acids described, which proceeds predominantly in the mitochondria. This Correlation with fatty acid metabolism represents a functional relationship of enoyl coenzyme A hydratase with FABP-3 and E-FABP. These are also fatty acid-binding proteins in their abundance in cancerous tissue changed in a characteristic way. in this connection FABP-3 is downregulated about 2.5-fold and E-FABP about 2.3-fold upregulated. Besides the connection with fatty acids was for FABP-3 a role in cell cycle control has been described (Seidita G. et al, 2000, Carcinogenesis 21: 2203-10). E-FABP has already been related described with various types of cancer and could be found in urine of cancer patients (Brouard M.C. et al., 2002, Melanoma Research 12: 627-31). The one observed by the inventors increased Abundance of this protein makes it especially in combination with the other two markers mitochondrial enoyl-coenzyme A hydratase and / or FABP-3 as a diagnostic marker for cancer in a special way suitable, since here between these different proteins a functional Context exists. It can in each case one of these proteins or particularly advantageously two or three of these proteins in their abundance compared with controls be observed, so that by the characteristic upregulation or downregulation of these proteins Conclusions on the presence of cancerous tissue can be made. The connection with the fatty acid metabolism also applies to more of the proteins described here, as explained below becomes.

Farther comprises the invention uses the protein serum amyloid P component (SAP) as a diagnostic Markers for Cancer. The inventors were able to show that this protein is in cancerous tissue is about 2.7 times lower in its frequency. The Investigation of downregulation of SAP compared with controls is therefore a characteristic feature of cancerous tissue suitable. SAP is a lectin-like acute phase protein (results from mice) the pentraxin family. The pentraxins are inducible by cytokines and increase in their concentration in the blood in infections or Trauma dramatically. So they play a role in the immune system.

Farther comprises the invention uses the protein 14-3-3 protein tau as diagnostic marker for Cancer. This protein is known to be involved in apoptotic processes. Furthermore, this protein has already been linked to Cancer described. However, an anti-oncogenicity was found (He H., 1997, Gan-To-Kagaku-Ryoho 24: 1448-53). In contrast, could Surprisingly, by the inventors an elevated one Level (1.8-fold) of 14-3-3 protein tau detected in cancerous tissue become. Invention according to therefore becomes a down-regulation of the protein in comparison with controls as characteristic Characteristic for Examined cancerous tissue.

Farther comprises the invention the use of the protein nuclear chloride ion channel protein as a diagnostic marker for Cancer, especially for Prostate cancer. In this regard, The inventors were able to increase the abundance of this protein by about 1.5-fold in cancerous tissues compared with controls. Thus, will preferably a downregulation of this protein by comparison examined with controls as a characteristic feature of cancerous tissue. This intracellular Anionic channel has already been associated with cell division and apoptosis (Ashley R.H., 2003, Mol. Membr. Biol. 20: 1-11).

Farther comprises the invention the use of the protein HES1 as a diagnostic Marker for cancer. The inventors were able to show that this protein is in Cancer tissue in its abundance about fourfold compared with controls elevated is. The T-test probability for a corresponding down-regulation was with p <0.0001 calculated. It is therefore preferable to downregulate this Protein compared with controls as a diagnostic trait for one Cancer disease examined. This protein is a specific splice variant (HES1 / Kpn-Ia) whose function is unknown so far. It contains one DJ1 PfPl domain and a mitochondrial localization is assumed. This Protein is expressed in many human tissues. The inventors For the first time, this protein was linked to cancer demonstrate.

Farther comprises the invention uses the proteasome alpha 2 subunit as a diagnostic marker for Cancer. Again, the inventors were able to connect for the first time detect this protein with cancers. there has been a about two-fold increase the abundance in cancerous tissue compared with controls. The T-test probabilities yielded for this protein has a value of p <0.009. Preferably, upregulation of the proteasome alpha 2 subunit compared with controls. Proteasomes are known to be at Processing of peptide presentation of the immune system over the MHC class I system involved, which in connection with the activity of killer T cells.

Farther comprises the invention the use of the protein adenine phosphoribosyltransferase as a diagnostic Markers for Cancer, especially for Prostate cancer. This protein has been used several times in the Related to cancer discussed. For example, it is described that this Protein downregulated in lymphocytes of breast cancer patients is. Furthermore, an overexpression of the protein has already been reported observed in colorectal carcinoma. The inventors could now show that this Protein in prostate cancer tissue about twice in its frequency elevated is. The T-test probabilities for this differential expression gave a value of p <0.007. Therefore, according to the invention a downregulation of the protein compared with controls as characteristic feature for Examined prostate cancer tissue.

In addition, the invention encompasses the use of the inorganic pyrophosphatase protein as a diagnostic marker for cancer, in particular for prostate cancer. Upregulation of this protein in lung cancer and colorectal cancer has been demonstrated. The inventors have now been able to show that this protein in prostate cancer tissue is upregulated about 1.6-fold compared with controls. These results showed a T-test probability of p <0.005. The reaction catalyzed by inorganic pyrophosphatase releases inorganic phosphate. This is related to the calcification processes involving annexins, particularly annexin A3, which are involved in the Ca ²⁺ flux. In particular, between the upregulation of annexin A3 and the downregulation of organic pyrophosphatase therefore has a functional relationship.

The various proteins mentioned as well as the proteins listed below can each for itself or in combination with other proteins to diagnostic Purposes are used.

Farther comprises the invention the use of at least one of the following Proteins as a diagnostic marker for cancer: ubiquitin-isopeptidase T, serum amyloid P-component (SAP), fatty acid-binding Protein 3 (FABP-3), galectin, microseminoprotein beta, heat shock protein 27 (HSP27), 14-3-3 protein beta, 14-3-3 protein zeta, nuclear chloride ion channel protein (CLIC-1), 14-3-3 protein tau, heat shock protein 90 (HSP90), protein disulfide isomerase (PDI), epidermal fatty acid-binding Protein (E-FABP), mitochondrial enoyl-coenzyme A hydratase, nucleophosmin, Annexin, in particular annexin A3, transgelin, triosephosphate isomerase, Aldolase A, HES1, proteasome alpha 2 subunit, adenine phosphoribosyltransferase and inorganic pyrophosphatase. It is particularly preferred if a down-regulation of at least one of the proteins ubiquitin isopeptidase T, serum amyloid P component (SAP), fatty acid binding protein 3 (FABP-3), Galectin, microseminoprotein beta, heat shock protein 27 (HSP27) or Transgelin is examined in comparison with controls. Farther it is preferable if in addition or alternatively an up-regulation of at least one of the following proteins in comparison with controls examined 14-3-3 protein beta, 14-3-3 protein zeta, nuclear chloride ion channel protein (CLIC-1), 14-3-3 protein tau, heat shock protein 90 (HSP90), protein disulfide isomerase (PDI), epidermal fatty acid-binding Protein (E-FABP), mitochondrial enoyl-coenzyme A hydratase, nucleophosmin, Annexin, in particular annexin A3, triosephosphate isomerase, aldolase A, HES1, proteasome alpha 2 subunit, adenine phosphoribosyltransferase and inorganic pyrophosphatase. It is particularly preferred if additionally downregulation of one or more of these proteins Annexin A2 is examined. Furthermore, it is particularly preferred that at least two proteins are examined.

In particularly advantageously at least two of the following Proteins used as diagnostic markers: ubiquitin isopeptidase T, heat shock protein 27 (HSP27), heat shock protein 90 (HSP90), protein disulfide isomerase (PDI), mitochondrial enoyl coenzyme A hydratase and / or nucleophosmin.

According to the invention could be shown that this different proteins in a characteristic way in cancerous tissue down-regulated or up-regulated compared to healthy tissue are. Details are given in the following table 1. This is the identification and quantification of the various Proteins in benign and malignant Tissue combined. The selection of proteins is based on a statistically significant differential expression of the proteins in benign (benign fraction) and malicious (Cancer fraction) tissue. The Accession Number corresponds to the respective one Number from the NCBI database. The theoretical molecular weight (MW) is the molecular weight calculated according to the database sequences has been. Under "scores" are those with help Understand the results of the MASCOT technique. The indication The PMF score refers to the Mowse score obtained from the MASCOT server In general, a PMF score higher than 65, a significant identifier is represented. The last two Columns summarize the quantification of protein spot intensities, the for the benign and malignant Tissue fractions were found.

Table 1:

In this context, will continue to the 5 referenced in tabular form the results of the protein spots with significant differential average expression in 21 patients along with different statistical values.

• 1. gi|1732411: Ubiquitin-Isopeptidase T; Isopeptidase T (isoT); ubiquitin specific protease 5; Ubiquitin carboxyl-terminal hydrolase 5; Ubiquitin thiolesterase 5; Ubiquitin-specific processing protease 5; Deubiquitinating enzyme 5; de-ubiquitinase.
• 2. gi|576259: Serum-Amyloid P-Komponente; Chain A; Serum Amyloid P Component (SAP).
• 3. gi|494781: Fettsäure-bindendes Protein 3 (FABP-3); Mammaryderived growth inhibitor (MDGI); fatty acid binding protein 3 (FABP-3); Heart-Type Fatty Acid Binding Protein (H-FABP); Muscle-Type Fatty Acid Binding Protein (M-Fabp).
• 4. gi|4504981: Galektin; galectin-1; 14 kDa beta-galactoside-binding lectin; beta galactoside soluble lectin; Beta-galactoside-binding lectin L-14-I; Galaptin; soluble galactoside-binding lectin; S-Lac lectin 1.
• 5. gi|225159: Mikroseminoprotein beta; beta-microseminoprotein; microseminoprotein beta; Immunoglobulin binding factor (IGBF); PN44; Prostate secreted seminal plasma protein; Prostate secretory protein of 94 amino acids (PSP-94); Seminal plasma beta-inhibin; seminal plasma protein.
• 6. nicht identifiziert
• 7. gi|662841: Hitzeschockprotein 27 (HSP27); heat shock protein 27; 27kDa heat shock protein 1 (HSP-27); Stress-responsive protein 27 (SRP27); Estrogen-regulated 24 kDa protein; 28 kDa heat shock protein.
• 8. gi|4507949: 14-3-3 Protein beta; 14-3-3 protein beta (14-3-3 beta); 14-3-3 protein alpha (14-3-3 alpha); Protein kinase C inhibitor protein-1; PKC inhibitor protein-1 (KCIP-1: auch 14-3-3 zeta); RNH-1.
• 9. gi|4507953: 14-3-3 Protein zeta; 14-3-3 zeta; 14-3-3 delta; KCIP-1 (auch 14-3-3 beta); YWHAZ; mitochondrial import stimulation factor S1 (MSF S1); Factor activating exoenzyme S; tryptophan monooxygenase activation protein zeta; tyrosine monooxygenase activation protein zeta.
• 10. gi|2073569: nukleäres Chloridionenkanal-Protein; chloride intracellular channel 1 (CLIC-1); nuclear chloride ion channel protein (p64CLCP); nuclear chloride channel; chloride channel ABP; Nuclear chloride ion channel 27 (NCC27); RNCC protein; Nuclear chloride ion channel 27 (NCC27).
• 11.: nicht identifiziert
• 12.: (Annexin A3, siehe 23.)
• 13. gi|5803227: 14-3-3 Protein tau; 14-3-3 tau; 14-3-3 theta; S15076 protein kinase regulator 14-3-3; HS1; tryptophan 5 -monooxygenase activation protein; tyrosine 3 -monooxygenase activation protein.
• 14. gi|13129150: Hitzeschockprotein 90 (HSP90); Heat shock protein 90 (HSP-90); Heat shock protein HSP 90-alpha; heat shock protein 90-alpha; 90 kDa heat-shock protein; heat shock protein 86 (HSP 86); Hspca; heat shock 90kDa protein 1; heat shock protein 1; Tumor specific transplantation 86 kDa antigen (TSTA).
• 15. gi|20070125: Protein-Disulfid-Isomerase (PDI); protein disulfide isomerase (PDI); Prolyl-4-hydroxylase beta; protein disulfide oxidoreductase; thyroid hormone binding protein p55; glutathione insulin transhydrogenase.
• 16. gi|4557581: epidermales Fettsäure-bindendes Protein (E-FABP); Fatty acid binding protein 5 (FABP-5); epidermal fatty acid-binding protein (E-FABP); Psoriasis-associated fatty acid-binding protein (PA-FABP); cutaneous fatty acid-binding protein (C-FABP); keratinocyte lipid binding protein (KLBP); DA11.
• 17. gi|12707570: mitochondriale Enoyl-Coenzym A-Hydratase; Mitochondrial Enoyl Coenzyme A hydratase; Mitochondrial enoyl-CoA hydratase; short-chain enoyl-CoA hydratase, mitochondrial; short-chain enoylcoenzyme A hydratase (SCEH).
• 18. gi|16307090: Nukleophosmin; Nucleophosmin; nucleolar phosphoprotein B23; Nucleolar protein NO38; numatrin; NPM(1).
• 19. gi|7768772: HES1 Protein, homolog zu E. coli and Zebrafisch ES1 Protein, anti-sigma cross-reacting protein homolog I alpha precursor, KNP-Ia/Kpn-I alpha, GT335, ähnlich zu E. coli SCRP27A and zu Zebrafisch ES1 [Homo sapiens].
• 20. gi|4506181: Proteasomen alpha 2-Untereinheit; proteasome subunit HC3, proteasome component C3; macropain subunit C3; multicatalytic endopeptidase complex subunit C3 [Homo sapiens].
• 21. gi|4502171: Adenine-Phosphoribosyltransferase; AMP pyrophosphorylase; AMP diphosphorylase; transphosphoribosidase.
• 22. gi|11056044: anorganische Pyrophosphatase; cytosolic inorganic pyrophosphatase; inorganic pyrophosphatase 1; pyrophosphate phospho-hydrolase [Homo sapiens].

The following is a compilation of English-language synonyms for these various Proteins listed. Here, the prefixed number corresponds to the numbering in Table 1. The first place is the German name of these proteins chosen in the rest of the description.

• 1. gi | 1732411: ubiquitin isopeptidase T; Isopeptidase T (isoT); ubiquitin specific protease 5; Ubiquitin carboxyl-terminal hydrolase 5; Ubiquitin thiolesterase 5; Ubiquitin-specific processing protease 5; Deubiquitinating enzyme 5; de-ubiquitinase.
• 2. gi | 576259: serum amyloid P component; Chain A; Serum Amyloid P Component (SAP).
• 3. gi | 494781: fatty acid-binding protein 3 (FABP-3); Mammaryderived growth inhibitor (MDGI); fatty acid binding protein 3 (FABP-3); Heart-Type Fatty Acid Binding Protein (H-FABP); Muscle-Type Fatty Acid Binding Protein (M-Fabp).
• 4. gi | 4504981: galectin; galectin-1; 14 kDa beta-galactoside-binding lectin; beta galactoside soluble lectin; Beta-galactoside-binding lectin L-14-I; Galaptin; soluble galactoside-binding lectin; S-Lac lectin 1.
• 5. gi | 225159: microseminoprotein beta; beta-microseminoprotein; microseminoprotein beta; Immunoglobulin binding factor (IGBF); Pn44; Prostate secreted seminal plasma protein; Prostate secretory protein of 94 amino acids (PSP-94); Seminal plasma beta-inhibin; seminal plasma protein.
• 6. not identified
• 7. gi | 662841: heat shock protein 27 (HSP27); heat shock protein 27; 27kDa heat shock protein 1 (HSP-27); Stress-responsive protein 27 (SRP27); Estrogen-regulated 24 kDa protein; 28 kDa heat shock protein.
• 8. gi | 4507949: 14-3-3 protein beta; 14-3-3 protein beta (14-3-3 beta); 14-3-3 protein alpha (14-3-3 alpha); Protein kinase C inhibitor protein-1; PKC inhibitor protein-1 (KCIP-1: also 14-3-3 zeta); RNH -1.
• 9. gi | 4507953: 14-3-3 protein zeta; 14-3-3 zeta; 14-3-3 delta; KCIP-1 (also 14-3-3 beta); YWHAZ; mitochondrial import stimulation factor S1 (MSF S1); Factor activating exoenzymes S; tryptophan monooxygenase activation protein zeta; tyrosine monooxygenase activating protein zeta.
• 10. gi | 2073569: nuclear chloride ion channel protein; chloride intracellular channel 1 (CLIC-1); nuclear chloride ion channel protein (p64CLCP); nuclear chloride channel; chloride channel ABP; Nuclear chloride ion channel 27 (NCC27); RNCC protein; Nuclear chloride ion channel 27 (NCC27).
• 11 .: not identified
• 12th: (Annexin A3, see 23.)
• 13. gi | 5803227: 14-3-3 protein tau; 14-3-3 dew; 14-3-3 theta; S15076 protein kinase regulator 14-3-3; HS1; tryptophan 5-monooxygenase activation protein; tyrosine 3-monooxygenase activation protein.
• 14. gi | 13129150: heat shock protein 90 (HSP90); Heat Shock Protein 90 (HSP-90); Heat Shock Protein HSP 90-alpha; heat shock protein 90-alpha; 90 kDa heat-shock protein; heat shock protein 86 (HSP 86); Hspca; heat shock 90kDa protein 1; heat shock protein 1; Tumor specific transplantation 86 kDa antigen (TSTA).
• 15. gi | 20070125: protein disulfide isomerase (PDI); protein disulfide isomerase (PDI); Prolyl 4-hydroxylase beta; protein disulfide oxidoreductase; thyroid hormone binding protein p55; glutathione insulin transhydrogenase.
• 16. gi | 4557581: epidermal fatty acid-binding protein (E-FABP); Fatty acid binding protein 5 (FABP-5); epidermal fatty acid-binding protein (E-FABP); Psoriasis-associated fatty acid-binding protein (PA-FABP); cutaneous fatty acid-binding protein (C-FABP); keratinocyte lipid binding protein (KLBP); DA11.
• 17. gi | 12707570: mitochondrial enoyl-coenzyme A hydratase; Mitochondrial Enoyl Coenzyme A hydratase; Mitochondrial enoyl-CoA hydratase; short-chain enoyl-CoA hydratase, mitochondrial; short-chain enoylcoenzyme A hydratase (SCEH).
• 18. gi | 16307090: nucleophosmin; Nucleophosmin; nucleolar phosphoprotein B23; Nucleolar protein NO38; numatrin; NPM (1).
• 19. gi | 7768772: HES1 protein homologous to E. coli and zebrafish ES1 protein, anti-sigma cross-reacting protein homologue I alpha precursor, KNP-Ia / Kpn-I alpha, GT335, similar to E. coli SCRP27A and to Zebrafish ES1 [Homo sapiens].
• 20. gi | 4506181: proteasomes alpha 2 subunit; proteasome subunit HC3, proteasome component C3; macropain subunit C3; multicatalytic endopeptidase complex subunit C3 [Homo sapiens].
• 21. gi | 4502171: adenine phosphoribosyltransferase; AMP pyrophosphorylase; AMP diphosphorylase; transphosphoribosidase.
• 22. gi | 11056044: inorganic pyrophosphatase; cytosolic inorganic pyrophosphatase; inorganic pyrophosphatase 1; pyrophosphate phospho-hydrolase [Homo sapiens].

Furthermore four more proteins could be identified, each one in cancerous tissue compared with control tissue in certain patient groups (Cluster analysis) upregulated or downregulated. In fact these are the proteins annexin A3, transgelin, Triosephosphate isomerase and aldolase A. Annexin A3 is expressed in cancerous tissue upregulated about 5-fold and transgelin is downregulated about 5-fold. Triosephosphate isomerase and Aldolase A are each expressed in cancerous tissue upregulated by about 20% or about 10%.

In this context, on the 3 which graphically illustrates the results of the cluster analysis. This results in the upregulation or downregulation of various proteins in cancerous tissues of certain patient groups or specific clusters, each of which is represented by a circle, in comparison with healthy tissue.

• 23. gi|4826643: Annexin A3; Annexin III; lipocortin III; anticoagulant protein III; Placental anticoagulant protein III (PAP III); 35 alpha calcimedin.
• 24. gi|4507359: Transgelin; SM22-alpha smooth muscle protein, 22 kDa actin-binding protein, Smooth muscle 22 protein, Actin-associated protein p27, 25 kDa F-actin-binding protein.

The English-language synonyms for annexin A3 and transgelin are as follows:

• 23. gi | 4826643: Annexin A3; Annexin III; lipocortin III; anticoagulant protein III; Placental anticoagulant protein III (PAP III); 35 alpha calcimedin.
• 24. gi | 4507359: Transgelin; SM22-alpha smooth muscle protein, 22 kDa actin-binding protein, smooth muscle 22 protein, actin-associated protein p27, 25 kDa F-actin-binding protein.

Additionally were yet more proteins identified compared with cancerous tissue Abnormal abundance with control tissue in certain patient groups showed or were up- or down regulated. With these proteins is ATP synthase, biliverdin reductase B, glucose-regulated protein, Prolyl 4-hydroxylase beta and dnak-like molecular chaperone. The ATP synthase is down-regulated and the rest of these Proteins are upregulated.

Interestingly, Many of the identified proteins have a relation to lipid metabolism. For annexin A3 and SAP were described as binding directly to lipids. Both Proteins are involved in phagocytic processes. With FABP-3 and E-FABP were two fatty acid-binding Proteins identified. Mitochondrial enoyl-coenzyme A hydratase is involved in β-oxidation of fatty acids involved. The activity of HSP27 is due to the activity the protein kinase C stimulates itself through the activity of phospholipases affected which convert phospholipids. HSP90 also affects phospholipid metabolism, there is an inhibition of HSP90 in an altered phospholipid metabolism results (Chung Y.L. et al., 2003, J. Natl. Cancer Inst. 95: 1624-33). Furthermore, PDI is probably also associated with lipid metabolism connected, since PDI acts as a multifunctional protein which i.a. at the Triglyceride transfer is involved (Horiuchi R. and Yamauchi K., 1994, Nippon-Rinsho 52: 890-5). In addition, inhibit the 14-3-3 Proteins the activity the protein kinase C and contain conserved sequences that the Pseudosubstrate domain of the protein kinase C and the C-terminus of the annexins. This all points to it There is a functional relationship between these different ones Proteins exists.

With the invention used diagnostic markers can detected various types of tumors and cancers become. In a particularly preferred embodiment of the invention acts the cancer to be diagnosed is prostate cancer, in particular a prostate carcinoma. As already mentioned, ask Prostate cancers are the most common malignant tumors in men Only if a prostate tumor is detected at an early stage can, a preventative surgical removal of the prostate can be be promising therapy. For the advanced, not More on an organ-limited disease is a preventive withdrawal the prostate is no longer sufficient. For these partially inoperative Prostate tumors may be considered an inhibition of male sex hormones to be pulled. Such an inhibition, preferably in combination with a surgical or pharmacological castration in part the proliferation and metastasis of the tumor and allowed thus its control for a certain period of time. However, most prostate tumors develop over time some resistance to this endocrinological Therapy. Other treatment options, such as The use of cytotoxic agents, gene therapy or immunotherapy are in clinical trials, could so far unfortunately still no resounding success. This it requires that one Prostate cancer as possible in a very early age Stage is detected, so that then can still be surgically treated with success. The described, according to the invention diagnostic Marker proteins are great Advantage for early detection suitable for prostate cancer.

In a particularly preferred embodiment of the use according to the invention, a specific subspecies of cancer, in particular a sub-type of prostate cancer, can be diagnosed by examining preferably several of the proteins mentioned. The inventors were able to show that in a so-called cluster analysis certain protein patterns reflect a characteristic upregulation or downregulation of different proteins which correlate with certain patient groups. In this case, the patients of a group each have a specific subtype of cancer, in particular of prostate cancer. According to the invention, it is therefore provided that by analyzing certain protein patterns, the patient can be assigned to a specific patient group or sub-type of cancer, so that this particular subtype of the cancer can be specifically treated with particular advantage. In this case, advantageously, certain combinations of proteins are analyzed. It is in this regard on the 3 which graphically represents the protein patterns that characterize the various patient groups are table. 4 shows in tabular form a summary of the protein patterns representing the different patient groups or subtypes of prostate cancer.

• Unterart a: heraufreguliertes Transgelin, stark herabreguliertes Galektin, stark herabreguliertes Mikroseminoprotein beta, herabreguliertes Fettsäure-bindendes Protein 3, keine oder geringe Veränderungen von epidermalem Fettsäure-bindenden Protein, keine oder geringe Veränderungen des nukleären Chloridionenkanal-Proteins, keine oder geringe Veränderungen des 14-3-3 Proteins beta, keine oder geringe Veränderungen des 14-3-3 Proteins zeta, keine oder geringe Veränderungen des 14-3-3 Proteins tau, keine oder geringe Veränderungen der Aldolase A, keine oder geringe Veränderungen der Serum-Amyloid P-Komponente, keine oder geringe Veränderungen der Triosephosphat-Isomerase und/oder keine oder geringe Veränderungen von Annexin A3.
• Unterart b: stark heraufregulierte Protein-Disulfid-Isomerase, stark heraufreguliertes Hitzeschockprotein 90, stark herabregulierte Ubiquitin-Isopeptidase T, heraufreguliertes 14-3-3 Protein beta, heraufreguliertes 14-3-3 Protein zeta, heraufreguliertes 14-3-3 Protein tau, heraufregulierte Aldolase A, heraufregulierte Triosephosphat-Isomerase, heraufreguliertes Annexin A3, herabreguliertes Transgelin, herabreguliertes Galektin, herabreguliertes Mikroseminoprotein beta, herabregulierte Serum-Amyloid P-Komponente, keine oder geringe Veränderungen des Fettsäure-bindenden Proteins 3 und/oder keine oder geringe Veränderungen des nukleären Chloridionenkanal-Proteins.
• Unterart c: stark heraufreguliertes nukleäres Chloridionenkanal-Protein, herunterregulierte Serum-Amyloid P-Komponente, keine oder geringe Veränderungen des Fettsäure-bindenden Proteins 3, keine oder geringe Veränderungen des 14-3-3 Proteins beta, keine oder geringe Veränderungen des 14-3-3 Proteins zeta, keine oder geringe Veränderungen des 14-3-3 Proteins tau, keine oder geringe Veränderungen der Aldolase A, keine oder geringe Veränderungen der Triosephosphat-Isomerase, keine oder geringe Veränderungen von Annexin A3, keine oder geringe Veränderungen des epidermalen Fettsäure-bindenden Proteins, keine oder geringe Veränderungen von Mikroseminoprotein beta, keine oder geringe Veränderungen von Galektin und/oder keine oder geringe Veränderungen von Transgelin.

For a diagnosis of the various subtypes of prostate cancer, a combination of different proteins is advantageously investigated in their abundance. Initially, at least one of the following proteins is analyzed as a general cancer marker: upregulated nucleophosmin, upregulated protein disulfide isomerase, up-regulated heat shock protein 90, up-regulated mitochondrial enoyl-coenzyme A hydrate, down-regulated heat shock protein 27 and / or down-regulated ubiquitin isopeptidase T. These proteins are analyzed in combination with at least one of the following proteins for the three subtypes of prostate cancer.

• Subspecies a: upregulated transglinine, highly downregulated galectin, highly downregulated microseminoprotein beta, downregulated fatty acid binding protein 3, no or minor alterations of epidermal fatty acid binding protein, no or minor changes in nuclear chloride channel protein, no or minor changes in 14- 3-3 protein beta, no or minor alterations of the 14-3-3 protein zeta, no or minor alterations of the 14-3-3 protein tau, no or minor alterations of aldolase A, no or minor changes of the serum amyloid P- Component, no or minor changes in triosephosphate isomerase and / or no or minor changes in annexin A3.
• Subspecies b: highly upregulated protein disulfide isomerase, highly upregulated heat shock protein 90, highly downregulated ubiquitin isopeptidase T, upregulated 14-3-3 protein beta, upregulated 14-3-3 protein zeta, upregulated 14-3-3 protein tau, up-regulated aldolase A, upregulated triosephosphate isomerase, upregulated A3 annexin, downregulated transgelin, downregulated galectin, down-regulated microseminoprotein beta, down-regulated serum amyloid P component, no or minor changes in fatty acid-binding protein 3 and / or no or minor changes in nuclear chloride ion channel protein.
• Subtype c: highly upregulated nuclear chloride ion channel protein, down-regulated serum amyloid P component, no or minor changes in fatty acid-binding protein 3, no or minor changes in 14-3-3 protein beta, no or minor changes in 14-3 -3 protein zeta, no or minor changes in the 14-3-3 protein tau, no or minor changes in aldolase A, no or slight changes in triosephosphate isomerase, no or minor changes in annexin A3, no or minor changes in epidermal fatty acid -binding protein, no or minor changes of microseminoprotein beta, no or minor changes of galectin and / or no or minor changes of transgelin.

at the use according to the invention The described proteins as diagnostic markers can be different Methods used to determine the frequency or abundance of Proteins in cancer tissue (or tissue to be examined) in comparison to analyze with control tissue. It is particularly advantageous if the proteins of the sample to be examined and the control sample gelelektrophoretisch be separated, for example, on a conventional Polyacrylamide gel. Subsequently is the abundance of the respective proteins in sample and control compared to each other. Because of the required resolution especially preferred are two-dimensional gels. on the other hand it is also possible pre-cleaning before the gel electrophoretic separation, so that too for example, with one-dimensional polyacrylamide gel electrophoresis sufficient separation and analyzability is achieved. Other methods for the separation of proteins can with Be used advantage, for example, conventional chromatographic methods, in particular column chromatographic Methods. It is particularly advantageous if the to be examined Sample and the control sample marked in different ways is, for example, by different isotopes. This will a comparison of the sample to be examined with the control in relation facilitated to the abundance of the respective proteins. In another preferred embodiment the proteins to be analyzed are analyzed by mass spectrometry, to allow accurate identification of the proteins.

In a further preferred embodiment the proteins to be examined are qualitatively using antibodies and quantitatively characterized. Here, polyclonal and / or monoclonal antibody be used against the particular proteins to be examined, which are used as diagnostic markers are directed. To the Qualitative and above all quantitative detection can be carried out using conventional immunoassays used, such as conventional enzyme-linked immunosorbent assays (ELISA). Furthermore you can also immunohistochemical methods and / or protein chips used become. For the Proof can for example, body fluids or tumor tissue. Especially for the proof of Annexin A3 are antibodies suitable.

In a further preferred embodiment the qualitative and quantitative proof of the diagnostic Marker proteins using oligonucleotides, for example in the course of a usual Polymerase chain reaction (PCR) can be used. in this connection it is a molecular genetic method in which selective certain DNA sections or RNA sections are amplified. hereby can be a qualitative and quantitative proof of the examined Proteins take place at the DNA or RNA level. Furthermore, with suitable oligonucleotides so-called hybridization experiments for example, conventional Northern or Southern blots carried out which also provides qualitative and quantitative statements can be made to the proteins at the RNA or DNA level. Of the The advantage of these detection methods with oligonucleotides lies in the good automation of these procedures. On the other hand must also be considered be that by this not necessarily Statements about the actual abundance of the protein are made, but merely Statements about the frequency the corresponding DNA or RNA sections. In this case, so in advance clarified whether the characteristic down- or down-regulation of the respective diagnostic marker proteins, for example at mRNA level, or whether the regulation is downstream of one of the transcription Level is accomplished.

The determined according to the invention characteristic change the abundance of the various proteins over control tissues affects especially on the activity of the respective proteins, for example on their enzymatic activity, out. Therefore, it is further preferred that in addition to the study of the Abundance or, alternatively, determines the activity of the respective proteins is compared and compared with controls. This too is under the To understand downregulation or downregulation of the different proteins. A corresponding investigation, for example, by conventional enzymatic Tests are done for the respective proteins to the expert. Furthermore, for example in the case of fatty acid-binding Proteins corresponding binding assays or comparable can be performed to so findings about the activity or the up or Downregulation of these proteins. The same applies to the others Proteins. For example, you can certain channel activities be measured to statements about the nuclear To make chloride ion channel protein. This may be for use the various proteins as diagnostic markers or the im utilized the following described diagnostic kit according to the invention become. Furthermore, such a measurement of the activity of the respective Proteins performed to test the effect of an active ingredient according to the invention for the production a drug is used to treat cancer as follows is described.

In a further preferred embodiment the use according to the invention be for an examination of the at least one protein exosomes from for example Patient material isolated and with respect to the protein or proteins analyzed. In particular, the protein pattern of one or more Examined proteins within the exosomes, so that hereby the diagnostically relevant up and / or downregulation of individuals or more proteins can be detected. A suitable Isolation of the exosomes from, for example, patient material can with usual Methods are carried out, which are familiar to the expert.

Farther comprises the invention provides a diagnostic kit containing at least one substance for proof of activity and / or expression of at least one of the described proteins comprises which are used as diagnostic markers as described above become. With the help of this diagnostic kit is preferably the activity and / or the expression of at least one of the following proteins detected: ubiquitin isopeptidase T, serum amyloid P component (SAP), nuclear chloride ion channel protein, mitochondrial enoyl-coenzyme A hydratase and / or annexin A3. Above all, such a diagnostic kit is used to to determine the respective abundance of at least one of these proteins, the opposite Controls in a characteristic way in cancerous tissue up or is down regulated. The abundance reflects here in the first place the expression of the protein. The diagnostic kit according to the invention is advantageously for the detection or screening of Cancers, in particular of prostate cancer, suitable In this way, in a particularly advantageous manner, an early detection of such diseases becomes. For example, with such a diagnostic kit or with the use according to the invention the described marker proteins also between benign or healthy and malicious Tissue can be distinguished, for example, between benign Prostatahyperplasia tissue and prostate cancer tissue. Preferably, such a diagnostic kit contains one or more antibodies or one or more oligonucleotides or oligonucleotide pairs, with one or more of the described proteins or the corresponding nucleic acids interact. With the help of these substances can thus qualitative and especially quantitative statements compared to these proteins to be taken with checks.

The samples to be examined and the control samples generally come from the body of a patient, for example, tissue samples or samples of body fluids, such as As blood, serum, lymph or urine, and processed in the familiar manner for a person skilled in the art. It is particularly advantageous if potentially malignant tissue, that is to say the tissue to be examined, and control tissue, ie benign tissue, are taken from the same patient and compared directly with one another. On the other hand, it is also possible to compare the abundance of the respective proteins with other standards, which are statistically determined, for example, from a large number of independent control samples. In the case of prostate cancer, benign and potentially malignant prostate tissue is advantageously taken from a patient undergoing prostatectomy. For example, the control tissue may be benign prostatic hyperplasia tissue.

The Invention a method of diagnosing cancer, wherein at least one the described proteins are analyzed in terms of their abundance and / or activity becomes. By the analysis of inventively ascertained down or Upregulation of these proteins in cancerous tissue can be made from these Findings Conclusions for the existence be pulled by cancerous tissue. Regarding further features of this inventive method Reference is made to the above description.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of annexin, in particular annexin A3, influenced, preferably inhibits, for the manufacture of a medicament for the treatment of cancer.

In a particularly preferred embodiment this use is at least the active ingredient a benzodiazepine derivative (Hofmann et al., 1998, J. Biol. Chem. 273 (5): 2885-2894). Especially preferred here is BDA250 (1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one), BDA452 (3- (R, S) - (L-Trypthophanyl) -1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one) and / or BDA753 (3- (R, S) -all-L- (NH-Trp-Gly-Tyr-Ala-H) -1,3-dihydro-1-methyl-5-phenyl-2H-1,4 benzodiazepin-2-one). Farther is also the use of diazepam (7-chloro-1,3-dihydro-1-methyl-5-phenyl-2H-1,4-benzodiazepin-2-one) prefers. Other derived from these substances molecules can with Advantage used according to the invention become. These are, in particular, those molecules which the activity of Block annexin A3.

In a special way, an annexin A3-specific antibody is also suitable as the active substance. These are preferably blocking antibodies and / or radiolabelled antibodies carrying, for example, ¹²⁵ l. With such antibodies can be carried out with advantage a so-called radio-immunotherapy, as is familiar to the expert.

In particularly preferably, with such agents, the activity and / or expression of annexin A3 in exosomes.

drugs the the activity and / or affect the expression of annexin A3, in particular inhibit, can continue to be advantageous for the manufacture of a medicament for treatment osteoarthritic cartilage degradation and / or arteriosclerotic lesions be used.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of the protein ubiquitin isopeptidase T and / or the activity and / or the expression of the protein disulfide isomerase (PDI), Preparation of a drug for the treatment of cancer. As already is the frequency or abundance of these proteins in cancerous tissue in more characteristic Way changed. In particular, the abundance of ubiquitin isopeptidase T is significantly decreased and Increases the abundance of protein disulfide isomerase (PDI). Influencing Expression or the activity of these Proteins towards the normal level, as realized in control fabrics, for example therefore a therapeutic approach to the treatment of cancer On the one hand, it therefore claims the use of an active substance, the activity and / or enhances the expression of ubiquitin isopeptidase T. To the Others claim the use of an active ingredient which is the substance of choice activity and / or inhibits the expression of the PDI. By such agents becomes the activity the mentioned Proteins to the usual Measure regulated, whereby a cancer can be effectively treated.

Further, the invention encompasses the use of at least one drug which affects the activity and / or expression of mitochondrial enoyl-coenzyme A hydratase for the manufacture of a medicament for the treatment of cancer. Preferably, this may be done in combination with an effect on the fatty acid-binding protein 3 (FABP-3) and / or the epidermal fatty acid binding protein (E-FAPB). It is preferred if an inhibiting agent for the activity and / or expression of mitochondrial enoyl-coenzyme A hydratase and / or E-FABP or an increasing active ingredient is used for the activity and / or expression of FABP-3.

Farther comprises the invention the use of at least one active ingredient, the the activity, Expression and / or localization of the serum amyloid P component (SAP) influenced, in particular boosts, for the manufacture of a drug for the treatment of cancer. It has been shown that the Localization of SAP may be altered in cancer. Therefore, can it is preferred according to the invention be that the lo calisation is influenced by SAP by using at least one active ingredient.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of the protein 14-3-3 protein tau affects, preferably inhibits, for the manufacture of a medicament for the treatment of cancer.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of the nuclear Chloride ion channel protein affects, preferably inhibits Preparation of a medicament for the treatment of cancer, in particular of prostate cancer.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of the protein HES1 affects, in particular inhibits, Preparation of a drug for the treatment of cancer.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of the proteasome alpha 2 subunit influenced, in particular inhibits, for the manufacture of a medicament for the treatment of cancer.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of the protein adenine phosphoribosyltransferase affected preferably inhibits, for the manufacture of a medicament for treatment of prostate cancer.

Farther comprises the invention the use of at least one active ingredient, the the activity and / or Expression of the protein anorgani cal pyrophosphatase influenced, in particular inhibits, for the manufacture of a medicament for the treatment of prostate cancer.

Furthermore comprises the invention the use of at least one active ingredient, the the activity and / or expression of at least one of the following proteins affected especially increases, for the manufacture of a medicament for treatment of cancer: ubiquitin-isopeptidase T, serum amyloid P component (SAP), fatty acid-binding Protein 3 (FABP-3), galectin, microseminoprotein beta, heat shock protein 27 (HSP27) and transgelin. About that included the invention the use of at least one active ingredient, the the activity and / or expression of at least one of the following proteins affected preferably inhibits, for the manufacture of a medicament for treatment of cancer: 14-3-3 protein beta, 14-3-3 protein zeta, nuclear chloride ion channel protein, 14-3-3 protein tau, heat shock protein 90 (HSP90), protein disulfide isomerase (PDI), epidermal fatty acid binding Protein (E-FABP), mitochondrial enoyl-coenzyme A hydratase, nucleophosmin, Annexin, in particular annexin A3, triosephosphate isomerase, aldolase A, HES1, proteasome alpha 2 subunit, adenine phosphoribosyltransferase and inorganic pyrophosphatase. It is particularly preferred if two or more drugs against at least two different proteins be combined with each other. In addition, it is preferable that in Combination with one or more of these agents another Active substance is used which controls the activity and / or expression of annexin A2 boosts.

at All of these proteins have been shown according to the invention to be in Characteristically in cancer tissue compared with control cells be up or down regulated. Therefore, it is claimed according to the invention these proteins in opposite ways by appropriate Downregulate active ingredients and so the activities, in particular the enzymatic activities, of achieving healthy control tissues. This can be different Cancers are treated successfully. Especially preferred Here is the production of drugs for the treatment of Prostate cancer.

The active ingredients used according to the invention may be, for example, peptides, proteins, so-called small molecular compounds or polynucleotides. These may in each case be known active substances which influence the activity and / or expression of the various proteins in a known manner, or they may be new active substances. These drugs can attack directly on the described proteins. On the other hand, it may also be preferred if these active substances Regulato ren, in particular activators or inhibitors, and / or biological precursors of these different proteins influence. Depending on whether an inhibition or an increase in the activity and / or the expression of the respective protein is preferred when influenced by the active ingredient, the active ingredient may be an agonist or an antagonist. As antagonists, for example, inhibiting antibodies come into question, which can reduce or block the activity of the respective protein. Furthermore, antibodies can be used in the course of a so-called radio-immunotherapy, wherein the antibodies radiolabel, z. B. ¹²⁵ I carry. Further examples of antagonistic active substances are deficient mutants or dominant negative mutants, which can be constructed by molecular biological methods. For example, they are enzymatically inactive, but compete with the particular substrate of the protein or enzyme to be inhibited, so that the activity of the protein is reduced as a result. Another example of an antagonistic agent is antisense molecules that can reduce the expression of a particular protein in a known manner. Substances which promote the expression of a particular gene or the conversion of mRNA into the active gene product can be used, for example, as agonistically active substances. These may be, for example, specific transcription factors or the like, which regulate the expression level of said proteins. In particular, small-molecule chemicals can be used with advantage.

to Increase in activity the described proteins, especially the ubiquitin isopeptidase T, FABP-3, SAP, galectin, microseminoprotein beta, HSP27 and Transgelin, for example, a component can be used, the one comparable or similar enzymatic activity developed. Furthermore, by an appropriate drug the activity the already existing enzyme molecules induced or increased become. On the other hand, it is also possible with suitable active ingredients the expression, ie the synthesis, corresponding enzymatic molecules to induce or increase. In this case, for example, the active ingredient also on certain precursor molecules, regulators, Activators or inhibitors of enzymes or other proteins attack.

Farther can also hormones or similar effective substances are used as active substances when they activity of the respective protein in the desired Influence way. So can For example, progesterone analogues are used to treat the mitochondrial Enoyl coenzyme A hydratase to inhibit.

In a particularly preferred embodiment the use according to the invention the active substance is at least one of the following Proteins themselves: ubiquitin-isopeptidase T, serum amyloid P-component, fatty acid-binding Protein 3, galectin, microseminoprotein beta, heat shock protein 27 and / or transgelin. In these proteins, the inventors show that this Proteins in cancerous tissues are lowered in their abundance, so that it is provided according to the invention is to increase their activity and / or expression of these To introduce proteins themselves as an active ingredient. This can happen to single of these proteins or preferably to combinations of Act different of these proteins. Furthermore, it can be provided be that parts of these proteins, for example peptides, or of the proteins derived molecules according to the invention as active ingredient be used.

In a particularly preferred embodiment the use according to the invention the agent (s) is provided in the form of exosomes or the administration of the active substance or substances by a Mediation over Exosomes. As a result, the immune response of the Patients affected be modulated, and in particular the T-cell response. exosomes are membrane vesicles, which are secreted in particular by hematopoietic cells become. As is known, induce exosomes derived from dendritic cells effective anti-tumor immune response, for example in mice.

By the use according to the invention a drug for the treatment of cancer is advantageously achieved that the Development or growth of a tumor diminished or inhibited is, and / or that one Metastasis of tumors is at least partially prevented or completely avoided.

Farther comprises the invention is a pharmaceutical composition containing at least an active ingredient according to the above Description and at least one pharmaceutically acceptable carrier. Regarding details to this pharmaceutical composition or to the active ingredient Reference is made to the above description. Suitable pharmaceutically acceptable carriers tap to the expert.

Furthermore, the invention also includes a method for the treatment of cancers, for example of prostate cancer, wherein at least one of the described active ingredients is administered. Bezüg Lich further features of this treatment method is made to the above description.

The Administration of the active substance can take place in various ways, for example, orally, intravenously, topically or by inhalation. Corresponding formulations are the Specialist familiar. The mode of administration depends especially on the disease to be treated as well as of course from the constitution of the patient. More details are available the expert.

Finally, the invention includes a method of searching for drugs for the treatment of cancer, in particular of prostate cancer, wherein in this method at least one protein is used, which is selected from the following group: Ubiquitin isopeptidase T, serum amyloid P component, fatty acid-binding Protein 3, galectin, microseminoprotein beta, heat shock protein 27, 14-3-3 protein beta, 14-3-3 protein zeta, nuclear chloride ion channel protein, 14-3-3 protein tau, heat shock protein 90, protein disulfide isomerase, epidermal fatty acid-binding Protein, Mitochondrial Enoyl Coenzyme A Hydratase, Nucleophosmin, Annexin, in particular annexin A3, transgelin, triosephosphate isomerase, Aldolase A, HES1, proteasome alpha 2 subunit, adenine phosphoribosyltransferase and inorganic pyrophosphatase. Particular preference is given here Ubiquitin isopeptidase T, serum amyloid P component (SAP), nuclear chloride ion channel protein, 14-3-3 protein tau, mitochondrial enoyl coenzyme A hydratase and / or annexin A3. Furthermore you can also offsprings or derivatives of these proteins are used, these being in particular homologous sequences or mutated forms of these proteins can act, for example, produced by molecular biology methods could be. Furthermore you can as descendants Parts of these proteins or subregions or combinations of the different Proteins and / or derivatives used become. The implementation of the method itself readily the skilled person, for example, a corresponding Protein or a derivative of which are expressed in a suitable expression system and using this system to interact with potential binding partners, especially inhibitors or activators. to Investigation of such interactions is for example suitable the two-hybrid system.

The described features and other features of the invention will be apparent from the following description of preferred embodiments in conjunction with the subclaims and the figures. Here you can the individual features for each be realized in or in combination with each other.

In show the pictures:

1 : Representation of a two-dimensional polyacrylamide gel with separated proteins. Isoelectric focusing was at pH 4-7. The numbered protein spots show those proteins that show aberrant abundance in cancerous and control tissues. The numbering corresponds to Table 1.

2 : Representation of a two-dimensional polyacrylamide gel with separated proteins. Isoelectric focusing was at pH 6-11. The numbered protein spots show those proteins that show aberrant abundance in cancerous and control tissues. The numbering corresponds to Table 1.

3 : Graphical representation of protein patterns characteristic of certain patient groups or subtypes of prostate cancer. Results with T-test probabilities of p <0.01 are shown in black, results with T-test probabilities of 0.01 <p <0.1 are shown in gray. Proteins that vary in expression within the different clusters are boxed.

4 : Tabular representation of the different protein levels of patient groups 1 to 3 compared to benign (healthy) and malignantly degenerated tissue. The data refer to percent of the protein spot size of cancerous tissue with standard error in relation to the total protein spot volume (benign + malignant). The T test result is represented as the probability that the distribution of the spot fractions from two given groups is significantly different. T-test results of at least 99% are in bold. Results below 95% are shown in light gray.

5 : Tabular list of protein spots with significant differential expression in all patients compared to benign (healthy) and malignant tissue based on the two-color ProteoTope analysis. "No. Obs. "Reflects the number of patients the spot was observed in. The spot fraction for benign (benign faction) and malignant (cancer fraction) with standard error is given as a percentage of the total spot volume (benign + malignant) -Test result is as a probability indicated that the distribution of the spot fractions for benign tissue deviated from the distribution found in cancerous tissues, taking into account all patients. The spots were selected on the assumption that the T-test probability is at least 99%.

to Identification of proteins relevant to the invention were Tissue samples from 23 patients were examined. cancerous tissue and control tissue was respectively processed and a two-dimensional Subjected to polyacrylamide gel electrophoresis (2D-PAGE). The isoelectric Focusing was at pH 4-7 and pH 6-11. The gel electrophoretic Results from two patients were unsuitable for further analysis. In another two patients, the results at pH 6-11 were unsatisfactory Results. Overall, therefore, the results of 21 patients in the pH range of 4-7 and the results of 19 patients in the pH range be evaluated by 6-11.

The Both different samples of each patient were each different Isotopes labeled, mixed and on a single two-dimensional Polyacrylamide gel separated by electrophoresis. The signals of each Isotops were subsequently added Detected separately, so that the protein pattern of the two Tissue samples could be compared directly (ProteoTope technology).

to Final identification of the proteins became analytical amounts (<1 μg) of the radiolabelled Proteins with preparative Quantities (> 200 μg) of the proteins the same sample together in preparative enrichment gels separated. The interesting protein spots were made from silver-stained preparative Gels excised, tryptically digested and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) (Bruker BiFlex or Ultra-Flex). Partially became an electrospray ionization ion trap mass spectrometry (ESI-MS) (Bruker Esquire).

On this way different proteins were clearly identified, which in matching Way an increased or a decreased abundance in cancerous tissue compared to control tissue exhibited.

at Some of the patient groups were formed in the analyzes, within which the abundances of the different proteins are studied were. In this so-called cluster analysis (Clustan Graphics 6.4), three groups of patients were identified, each of them characteristic Protein expression patterns showed. With the help of this procedure the proteins were annexin A3, transgelin, triosephosphate isomerase and Aldolase A identified for specific patient populations showed characteristic abundances.

tissue samples

healthy Prostate tissue and malignant prostate tissue was examined by patients who had previously undergone a prostatectomy. Patients were treated with the help of PSA (prostate specific antigen) Screening was examined, and the tumors were confirmed by ultrasound. A Consent of each patient was obtained prior to the operation.

Immediately after removal of the prostate this was transferred into a sterile box and Chilled there. There were 0.5 - 1 cm thick tissue sections made in a left and a right half were divided. These were embedded in a freezing matrix and shock-frozen. The rest of the prostate was fixed in formalin and according to usual Further processed standard procedure. For the production of tissue samples were thin Sections taken from both sides of the prostate and hematoxylin-eosin stained. These sections were stored at -80 ° C until use. Control tissue samples were taken from non-tumor-infested regions and an identical Subjected to treatment.

sample preparation

The Proteins were dissolved in 100 μl boiling 2% SDS, 0.1 M Tris, pH 8.8 and the protein concentration using the bicinchoninic acid method (BCA) determined.

The iodination with I-125 or I-131, the two-dimensional polyacrylamide gel electrophoresis and the data analysis were carried out by conventional methods (Cahill et al., 2003, Rapid Communications in Mass Spectrometry 17: 1283-1290). Radioactive iodine was from Amersham Bioscience (Freiburg). The protein iodination reactions with either I-125 or I-131 were each separately with identical iodine concentrations carried out.

Polyacrylamide gel electrophoresis

For the mission identical amounts of protein were labeled on the polyacrylamide gels Samples (cancer tissue and control tissue) mixed together. For the isoelectric Focusing (IEF) over the pH ranges 4-7 and 6-11 were the samples in a conventional Sample buffer on 18 cm immobilized pH gradient (IPG) strips (Amersham Bioscience). IEF as the first dimension of 2D PAGE protein separation was performed on a multiphor apparatus (Amersham Bioscience). The second dimension (SDS-PAGE) was performed on an ISO-DALT apparatus (Höfer). The Gels were dried, laminated to 80 micron plastic film, and subsequently was the radioactive measurement of the signals from both radioisotopes performed.

By the chosen one Analysis of the different radioisotopes could be a quantitative, multicolored differential representation of the proteins of different Samples are achieved. Therefore, a direct comparison of integrated protein spot intensities of the two samples, which were separated on a gel, for the further Analysis can be used. The analysis on a gel has the advantage that variations between two or more gels do not matter. The biggest potential Error source is a different stoichiometric mark with one or the other isotope. This was made by the manufacture excluded from gels with reversed markings. The is called so that the Control sample and the cancerous tissue sample each with the one and also marked with the other isotope and inverse each other were compared. Since the protein patterns matched the inverse labeling procedures, the qualitative criteria were met. With the help of computer-aided procedures The different isotope signals were each in a different one Color (blue or orange) made visible, so that consistent differences in the abundance between the samples in each case one or the other Color depending on the selected Isotope labeling appeared. For details, Cahill et al., 2003, Rapid Communications in Mass Spectrometry 17: 1283-1290.

image analysis

The Differential protein expression analysis is based on the reliable differential Quantification of protein spots on the described polyacrylamide gels. For one quantitative image analysis became the Phoretix 2D Advanced software (Nonlinear Dynamics) applied, with their own adjustments made were.

protein identification by mass spectrometry

It In principle, two mass spectrometric methods were used. On the one hand the very fast and reliable identification of high abundant proteins by peptide mass fingerprinting with MALDI-TOF MS. The identification of very low abundant proteins was with the more time-consuming LC-ESI-IonTrap-MS / MS or MALDI-TOF-TOF Method performed. Summarized were Gelstücke the chosen Cut out protein spots and carry the proteins in these gel pieces Help of trypsin digested. The resulting solution was initially with the peptide mass fingerprint procedure based on MALDI-TOF-MS analyzed. For the protein spots, for that in this way no clear identification was possible, was the slower fragment ion analysis based on MALDI-TOF-TOF or LC-ESI-IonTrap-MS / MS in addition carried out. A detailed description of these methods is Vogt et al., 2003, Rapid Communication in Mass Spectrometry 17: 1273-1282 and Vogt et al., 2003, Molecular Cellular Proteomics 2: 795.

identification the proteins

to Identification of the proteins were the peptide masses by mass spectrometry were obtained over the NCBI protein database evaluated. This was done with the help of the program MASCOT Version 1.9 (Matrix Science, London, UK).

quantitative image analysis

Quantitative analyzes were performed using the digital data recorded with a photomultiplier of a radio imager for each pixel of the image matrix. The protein spot limits were defined using the software Phoretix 2D Advanced (Nonlinear Dynamics) and the pixel values within the spot integrated region after subtraction of a suitable background signal. Based on the complete data generated, a detailed quantification of the detected protein spots was performed. Table 1 summarizes these results.

The 1 and 2 each show the positions of the selected protein spots, once at isoelectric focusing at pH 4-7 ( 1 ) and in the other case at pH 6-11 ( 2 ).

Claims (47)

Use of the protein annexin A3 as a diagnostic Brands for Cancer. Use according to claim 1, characterized that one Upregulation of Annexin A3 compared with controls studied becomes. Use according to claim 1 or claim 2, characterized characterized in that a Downregulation of annexin A3 in combination with downregulation Annexin A1, Annexin A2 and / or Annexin A5 is examined. Use of the protein ubiquitin isopeptidase T and / or protein disulfide isomerase (PDI) as a diagnostic marker for cancer. Use according to claim 4, characterized that one Downregulation of ubiquitin isopeptidase T and / or upregulation the protein disulfide isomerase (PDI) compared to controls becomes. Use of the protein mitochondrial enoyl coenzyme A-hydratase as a diagnostic marker for cancer. Use according to claim 6, characterized that one Up-regulation of mitochondrial enoyl-coenzyme A hydratase in the Comparison with controls is examined. Use of the protein serum amyloid P component (SAP) as a diagnostic marker for cancer. Use according to claim 8, characterized that one Downregulation of the serum amyloid P component (SAP) in comparison is examined with controls. Use of protein nuclear chloride ion channel protein as a diagnostic marker for Prostate cancer. Use according to claim 10, characterized that one Downregulation of the nuclear chloride ion channel protein is examined in comparison with controls. Use of the protein HES1 as a diagnostic Markers for Cancer. Use according to claim 12, characterized that one Downregulation of HES1 was examined in comparison with controls becomes. Use of the proteasome alpha 2 subunit as a diagnostic marker for Cancer. Use according to claim 14, characterized that one Upregulation of the proteasome alpha 2 subunit in comparison is examined with controls. Use of the protein adenine phosphoribosyltransferase as a diagnostic marker for Prostate cancer. Use according to claim 16, characterized that one Downregulation of adenine phosphoribosyltransferase compared is examined with controls. Use of the protein inorganic pyrophosphatase as a diagnostic marker for Prostate cancer. Use according to claim 18, characterized that one Upregulation of inorganic pyrophosphatase in comparison is examined with controls. Use of the proteins ubiquitin isopeptidase T and serum amyloid P component (SAP) as a diagnostic marker for cancer, preferably downregulation of the proteins compared to controls becomes. Use of at least two proteins that are selected from the group consisting of ubiquitin isopeptidase T, heat shock protein 27 (HSP27), heat shock protein 90 (HSP90), protein disulfide isomerase (PDI), mitochondrial enoyl coenzyme A hydratase and nucleophosmin as diagnostic Markers for cancer, wherein a down-regulation of ubiquitin isopeptidase T and / or Heat shock protein 27 (HSP27) and / or upregulation of Heat shock protein 90 (HSP90), protein disulfide isomerase (PDI), mitochondrial enoyl coenzyme A hydratase and / or nucleophosmin is examined in comparison with controls. Use according to one of the preceding claims, characterized characterized in that Cancer is prostate cancer. Use according to one of the preceding claims, characterized characterized in that the study of one or more proteins subspecies of cancer, especially from prostate cancer. Use according to claim 23, characterized that at least a protein according to claim 21 in combination with at least one protein selected from the group consisting of serum amyloid P component (SAP), fatty acid binding Protein 3 (FABP-3), galectin, microseminoprotein beta, 14-3-3 protein beta, 14-3-3 protein zeta, nuclear Chloride ion channel protein, 14-3-3 protein tau, epidermal fatty acid-binding Protein (E-FABP), annexin A3, transgelin, triosephosphate isomerase and aldolase A, be studied, with no or small changes of SAP, a downregulation of FABP-3, a strong down-regulation of galectin, a strong downregulation of microseminoprotein beta, no or small changes of 14-3-3 protein beta, no or small changes of 14-3-3 protein zeta, no or small changes of nuclear Chloride ion channel protein, no or small changes of 14-3-3 protein dew, no or little changes from E-FABP, no or minor changes from Annexin A3, one Upregulation of transgelin, no or small changes of triosephosphate isomerase and / or no or minor changes of aldolase A in comparison with controls. Use according to claim 23, characterized that at least a protein according to claim 21 in combination with at least one protein selected from the group consisting of serum amyloid P component (SAP), fatty acid binding Protein 3 (FABP-3), galectin, microseminoprotein beta, 14-3-3 protein beta, 14-3-3 protein zeta, nuclear Chloride ion channel protein, 14-3-3 protein tau, annexin A3, transgelin, Triosephosphate isomerase and aldolase A, being investigated a strong down-regulation of PDI, a strong down-regulation of HSP90, a strong down-regulation of ubiquitin-isopepti dase T, a down-regulation of SAP, no or little changes of FABP-3, a downregulation of galectin, a downregulation of microseminoprotein beta, a downregulation of 14-3-3 protein beta, a downregulation of 14-3-3 protein zeta, a downregulation of 14-3-3 protein tau, no or small changes of nuclear chloride ion channel protein, a downregulation of annexin A3, a downregulation of transgelin, upregulation of triosephosphate isomerase and / or a Upregulation of aldolase A compared with controls studied become. Use according to claim 23, characterized in that at least one protein according to claim 21 in combination with at least one protein selected from the group consisting of serum amyloid P component (SAP), fatty acid binding protein 3 (FABP-3), galectin, Microseminoprotein beta, 14-3-3 protein beta, 14-3-3 protein zeta, nuclear chloride ion channel protein, 14-3-3 protein tau, epidermal fatty acid binding protein (E-FABP), annexin A3, transgelin, triosephosphate Isomerase and aldolase A, being examined, showing a downregulation of SAP, no or small changes of FABP-3, no or small changes of galectin, no or small changes of microseminoprotein beta, no or small changes of 14-3-3 protein beta , no or small changes of 14-3-3 protein zeta, a strong upregulation of nuclear chloride channel protein, no or small changes of 14-3-3 protein tau, no or small change no or minor changes of transgenin, no or minor changes of triosephosphate isomerase and / or no or minor changes of aldolase A in comparison to be examined with controls. Use according to one of the preceding claims, characterized characterized in that at least one protein using polyacrylamide gel electrophoresis, in particular two-dimensional gel electrophoresis, mass spectrometry, antibodies ELISA, immunohistochemistry, protein chips and / or oligonucleotides, especially polymerase chain reaction (PCR). Use according to one of the preceding claims, characterized characterized in that for a study the at least one protein exosomes isolated and / or analyzed become. Diagnostic kit comprising at least one substance for proof of activity and / or expression of at least one protein selected from the group consisting of ubiquitin isopeptidase T, serum amyloid P component (SAP), nuclear Chloride ion channel protein, mitochondrial enoyl-coenzyme A hydratase and Annexin A3 for the detection of cancers, in particular of prostate cancer. Use of at least one active ingredient containing the activity and / or expression of the protein annexin A3, in particular inhibits, for the manufacture of a medicament for the treatment of cancer. Use according to claim 30, characterized that the Active ingredient at least one benzodiazepine derivative, in particular BDA250 and / or BDA452, is. Use according to claim 30 or claim 31, characterized characterized in that activity and / or expression of the protein annexin A3 in exosomes. Use of at least one active ingredient containing the activity and / or expression of the proteins ubiquitin isopeptidase T and protein disulfide isomerase (PDI) affected, for the manufacture of a medicament for the treatment of cancer, wherein preferably the active ingredient is the activity and / or expression of the ubiquitin isopeptidase T increases and / or the active substance the activity and / or expression of the protein disulfide isomerase (PDI) inhibits. Use of at least one active ingredient containing the activity and / or expression of the protein mitochondrial enoyl-coenzyme A hydratase affected for the manufacture of a medicament for the treatment of cancer. Use according to claim 34, characterized that the Active substance the activity and / or expression of the mitochondrial enoyl-coenzyme A hydratase inhibits. Use of at least one active ingredient containing the Activity, Expression and / or Localization of the Protein Serum Amyloid P Component (SAP) affected especially increases, for the manufacture of a medicament for treatment of cancer. Use of at least one active ingredient containing the activity and / or expression of the protein affects nuclear chloride channel protein, in particular inhibits, for the manufacture of a medicament for the treatment of prostate cancer. Use of at least one active ingredient containing the activity and / or expression of the protein HES1, in particular inhibits Preparation of a drug for the treatment of cancer. Use of at least one active ingredient containing the activity and / or expression of the proteasome alpha 2 subunit, in particular inhibits, for the manufacture of a medicament for the treatment of cancer. Use of at least one active ingredient containing the activity and / or expression of the protein adenine phosphoribosyltransferase affected especially for the manufacture of a medicament for treatment of prostate cancer. Use of at least one active ingredient containing the activity and / or expression of the protein affects inorganic pyrophosphatase, in particular inhibits, for the manufacture of a medicament for the treatment of prostate cancer. Use according to one of claims 30 to 41, characterized that the Cancer is prostate cancer. Use according to one of claims 30 to 42, characterized that the Active ingredient an agonist, antagonist, antibody, a deficient mutant, is a dominant negative mutant and / or an antisense molecule. Use according to one of claims 30 to 43, characterized that the Active ingredient is at least one protein selected from the group ubiquitin isopeptidase T, serum amyloid P-component (SAP), fatty acid-binding Protein 3 (FABP-3), galectin, microseminoprotein beta, heat shock protein 27 (HSP27) and transgelin. Use according to one of claims 30 to 44, characterized that the Drug is provided in the form of exosomes. A pharmaceutical composition comprising at least an active ingredient according to a the claims 30 to 45 and at least one pharmaceutically acceptable carrier. Method of searching for active ingredients for treatment of cancer, characterized in that at least one protein selected from the group consisting of ubiquitin isopeptidase T, serum amyloid P component (SAP), nuclear Chloride ion channel protein, 14-3-3 protein tau, mitochondrial enoyl coenzyme A hydratase, annexin A3, HES1, proteasome alpha 2 subunit, Adenine phosphoribosyltransferase and inorganic pyrophosphatase and / or at least one offspring thereof, is used.