WO1999013334A2 - Procedes de diagnostic des stades precancereux du foie et des reins - Google Patents

Procedes de diagnostic des stades precancereux du foie et des reins Download PDF

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Publication number
WO1999013334A2
WO1999013334A2 PCT/DE1998/002711 DE9802711W WO9913334A2 WO 1999013334 A2 WO1999013334 A2 WO 1999013334A2 DE 9802711 W DE9802711 W DE 9802711W WO 9913334 A2 WO9913334 A2 WO 9913334A2
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irs
antibody
fragment
reagent
liver
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PCT/DE1998/002711
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German (de)
English (en)
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WO1999013334A3 (fr
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Dirk Nehrbass
Fritz Klimek
Peter Bannasch
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Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts
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Publication of WO1999013334A2 publication Critical patent/WO1999013334A2/fr
Publication of WO1999013334A3 publication Critical patent/WO1999013334A3/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

Definitions

  • the present invention relates to methods for the diagnosis of early cancer precursors in the liver and kidneys, the presence of the insulin receptor substrate (IRS-1) in glycogenotic foci (GSF) and / or mixed-cell foci in a liver or kidney tissue sample (MCF) is detected on the basis of the binding of a specific reagent, for example an antibody, and the visualization of the complex formed.
  • the present invention further relates to the use of the method for testing substances that are suspected of being able to produce liver and / or kidney cancer.
  • Hepatocellular carcinoma is one of the most common malignant neoplasms in humans and causes at least 250,000 deaths worldwide every year.
  • Chronic infections with the hepatitis B or hepatitis C virus, the intake of foods that contain hepatocarcinogens, for example the naturally occurring aflatoxin B 1 f, and alcohol abuse are considered to be the main risk factors for the development of HCC.
  • an increased risk of developing hepatocellular adenoma has been observed in women taking contraceptives and in both women and men treated with anabolic steroid hormones.
  • liver cancer Other substances that occur in the human environment, it could be shown for at least in rodents that they are involved in the development of liver cancer, include at ⁇ play as nitrosamines, organic chlorinated pesticides, polychlorinated biphenyls le, phthalates and hypolipidaemia effecting Medication.
  • a decisive disadvantage of the diagnosis using these markers is, however, that either no distinction between early and late tissue changes in the development of cancer is possible, or that the tumor precursors cannot be clearly differentiated from the surrounding tissue for diagnostic purposes.
  • the latter applies in particular to the determination of glycogen, since glycogen is also present in large quantities in unchanged tissue. It is therefore not possible with the diagnostic method described in the prior art to detect early stages of cancer, which is an essential prerequisite for early and thus effective therapy.
  • the object of the present invention is therefore to provide a diagnostic method for diagnosing early precancerous stages in the liver and kidneys.
  • the diagnostic method according to the invention allows the detection of early stages of cancer in the liver and kidneys and is therefore suitable for early cancer detection. It is based on the fact that the presence of the insulin receptor substrate (IRS-1) in glycogenotic foci (GSF) and / or mixed cell foci (MCF) is detected in a tissue sample by binding to a reagent.
  • IRS-1 insulin receptor substrate
  • GSF glycogenotic foci
  • MCF mixed cell foci
  • Carcinogenesis in the liver and kidney is characterized in that first preneoplastic foci (glycogenotic foci, GSF; mixed cell foci, MCF; basophilic foci, BCF) develop from which hepatocellular carcinoma (HCC) or renal cell carcinoma (NCC) develops.
  • GSF glycogenotic foci
  • MCF mixed cell foci
  • BCF basophilic foci
  • GSF hepatocellular carcinoma
  • NCC renal cell carcinoma
  • the conversion of GSF to HCC or NCC leads to enzymatic changes in the liver, whereby glucose is preferably metabolized via the pentose phosphate cycle and glycolysis.
  • the glycogen originally stored in excess then gradually decreases, on the other hand there is an increase in basophilic ribosomes and cell proliferation.
  • the basophilic neoplastic phenotype finally emerges via the intermediate stage of the MCF.
  • the GSF metabolic pattern corresponds to the response of the hepatocytes to insulin.
  • Insulin is a pleiotropic hormone that affects the metabolism and mitosis in hepatocytes. This is done by a signal transduction cascade, in which IRS-1 is the intracellular substrate of insulin receptor tyrosine kinase.
  • IRS-1 is the intracellular substrate of insulin receptor tyrosine kinase.
  • the overexpression of IRS-1 in HCC or NCC could be demonstrated.
  • overexpression of IRS-1 in liver cell lines leads to cell transformation. So far, however, the expression of IRS-1 has not been investigated in the early stages of liver and renal cell cancer development.
  • IRS-1 is strongly overexpressed in the early, glycogen-storing cancer precursors (GSF, MCF), whereas it occurs only in very small, histochemically undetectable amounts in normal liver and kidney tissue.
  • GEF glycogen-storing cancer precursors
  • MCF glycogen-storing cancer precursors
  • expression of IRS-1 in more than 400 preneoplastic foci of altered hepatocytes (FAH) and in 1 2 hepatocellular carcinomas (HCC; induced with N-nitrosomorpholine, NNM) was induced in rats Western blots and examined by immunohistochemical detection. Using Western blots, IRS-1 in the liver of NNM-treated and untreated rats can be detected. However, IRS-1 was undetectable in normal liver parenchyma.
  • IRS-1 was overexpressed especially in the GSF and MCF, which had also stored glycogen excessively. It is therefore clear that overexpression of IRS-1 in GSF and MCF leads to preneoplastic glycogenosis and is an early event in carcinogenesis. During the progression of the early GSF to MCF, BCF and HCC or NCC, the expression of IRS-1 is then downregulated again. The preneoplastic insulinometric phenotype is gradually converted into the malignant neoplastic phenotype. This knowledge gained in the animal model also coincides with previous studies on humans, so that the above results are fully transferable. Thus, the overexpression of IRS-1 can be used as a specific marker for early cancer precursors in the liver and kidneys.
  • the present invention thus relates to methods for the diagnosis of early precancerous lesions in the liver and kidneys, which is characterized in that a liver or kidney tissue sample, or from nucleic ⁇ obtained ren with the insulin receptor substrate (IRS-1) brings specific reagent into contact and detects the presence of IRS-1 in glycogenetic foci (GSF) and / or mixed cell foci (MCF) based on the specific binding to the reagent.
  • a liver or kidney tissue sample or from nucleic ⁇ obtained ren with the insulin receptor substrate (IRS-1) brings specific reagent into contact and detects the presence of IRS-1 in glycogenetic foci (GSF) and / or mixed cell foci (MCF) based on the specific binding to the reagent.
  • GSF glycogenetic foci
  • MCF mixed cell foci
  • the detection methods which can be used for the diagnostic method according to the invention are known to the person skilled in the art and comprise immunohistochemical detection methods (for example the APAAP method) or the detection of IRS-1 by means of suitable nucleic acid probes, since the presence of overexpressed IRS-1 with a greatly increased concentration of the corresponding RNA is correlated.
  • the cDNA coding for IRS-1 was published by Sun et al., Nature 352, pp. 72-77 (1 991) and it is no problem for a person skilled in the art knowing this sequence to select the suitable probes for detecting the IRS-1 mRNA and construct. For this comes the in-situ known to the expert Hybridization, Northern Blot or Dot Blot in question.
  • the detection methods include taking a tissue sample from the liver, producing tissue sections, incubating the tissue sections with a reagent specific for IRS-1 and visualizing the specific complexes obtained in the tissue sections.
  • tissue sample of the liver can be removed by various methods known to the person skilled in the art, for example by means of fine needle biopsy or laparoscopic biopsy.
  • tissue sections are produced by customary methods, for example by means of a frozen section. The prepared tissue sections are then treated with the reagent specific for IRS-1.
  • Suitable reagents for the method according to the invention include IRS-1 specific antibodies and nucleic acid probes (DNA or RNA) hybridizing with IRS-1 mRNA.
  • the tissue sample is brought into contact with a polyclonal antibody, a monoclonal antibody or a fragment thereof, which can recognize IRS-1, as a reagent and so it is then determined whether the antibody or the fragment thereof is attached to the tissue sample is bound.
  • a suitable polyclonal antibody is, for example, an antibody directed against IRS-1, as is available from Upstate Biotechnology Inc., Lake Placid, NY, USA.
  • a monoclonal antibody is preferably used in the method according to the invention. used or a fragment of it.
  • Methods for producing monoclonal antibodies are also known to the person skilled in the art. For this purpose, for example, Zeil hybrids are produced and cloned from antibody-producing cells and bone marrow tumor cells. A clone is then selected which produces an antibody which is specific for IRS-1. This antibody is then made. Examples of cells that produce antibodies are spleen cells, lymph node cells, B-lymphocytes, etc. Examples of animals that can be immunized for this purpose are mice, rats, horses, goats and rabbits. The myeloma cells can be obtained from mice, rats, humans or other sources.
  • Cell fusion can be carried out, for example, by the well-known Köhler and Milstein method.
  • the hybridomas obtained by cell fusion are screened using IRS-1 according to the enzyme-antibody method or a similar method.
  • clones are obtained using the limit dilution method.
  • the clones obtained are implanted intraperitoneally in BALB / c mice, the ascites are removed from the mouse after 10 to 14 days, and the monoclonal antibody is purified by known methods (for example ammonium sulfate fractionation, PEG fractionation, ion exchange chromatography, gel chromatography or affinity chromatography).
  • the antibody obtained can be used directly or a fragment thereof can be used.
  • fragment means all parts of the monoclonal antibody (for example Fab, Fv or “single chain Fv” fragments) which have the same epitope specificity as the complete antibody.
  • the monoclonal antibody mentioned is an antibody derived from an animal (e.g. mouse), a humanized antibody or a human antibody or a fragment thereof.
  • tissue section After treatment of the tissue section with the antibody, further treatment is carried out to visualize and localize the immunohistochemical reaction using special reagents which are labeled with certain enzymes, fluorescent dyes or radioactive compounds.
  • special reagents which are labeled with certain enzymes, fluorescent dyes or radioactive compounds.
  • the evaluation of the Specifically stained tissue sections are preferably made visually using microscopic magnification.
  • GSF are characterized by an excessive storage of glycogen, which is shown on the alcohol-fixed tissue section by treatment with the Periodod-Schiff's reagent as red granules in the cytoplasm of the liver cells.
  • the MCF are characterized by a mixture of red-stained glycogen storage cells and cells with less glycogen, which in extreme cases no longer contain any glycogen and then by counter-staining of the sections with basic dyes, e.g. toluidine blue or cuprolin blue, which stain the ribosomes in the cytoplasm, as blue cells in Appearance.
  • a nucleic acid probe hybridizing with the IRS-1 mRNA is used as the reagent which detects IRS-1.
  • the detection is carried out via the in-situ hybridization known to the person skilled in the art, Northern blot or dot blot. These methods are professional rea ⁇ accordingly known and it is also familiar to win him from the tissue sections mRNA.
  • the complexes formed between IRS-1 and reagent can be made visible by using a differently labeled reagent or other substrates.
  • suitable labels are enzymes (peroxidase, galactosidase), fluorescent (fluorescein isothiocyanate, rhodamine isothiocyanate) or radioactive compounds.
  • enzymes peroxidase, galactosidase
  • fluorescent fluorescein isothiocyanate
  • rhodamine isothiocyanate fluorescein isothiocyanate
  • radioactive compounds radioactive compounds.
  • colored substrates bromo-chloro-indolyl phosphate / nitro-blue tetrazolium, diaminobenzidine, aminoethyl carbazole
  • luminescent compounds dioxetanes
  • the present invention further relates to the use of the method for determining substances which can induce liver and / or kidney cancer.
  • NEN taking advantage of the observations described above, ie the substance to be examined is applied to an animal, after a suitable period of time a tissue sample is taken, which is in contact with a reagent specific for the insulin receptor substrate (IRS-1) brought and the possible presence of IRS-1 in glycogenotic foci (GSF) and / or mixed cell foci (MCF) detected based on the specific binding to the reagent.
  • GSF glycogenotic foci
  • MCF mixed cell foci
  • the substance to be examined is applied to the animal, for example a mouse, a rabbit or a rat, in a suitable manner and in suitable concentrations.
  • a dose of 10% of the dose at which 50% of the test animals die after a single administration (L D 50) is preferred, possibly after sensitization of the liver by one administration of a known carcinogen (eg diethylnitrosamine).
  • a known carcinogen eg diethylnitrosamine.
  • fine needle biopsies are carried out or tissue samples are taken from the killed animals and the tissue sections are examined for the expression of IRS-1 in GSF and MCF.
  • the inventors surprisingly found that glycogenotic foci also appeared in the kidney long before kidney cell carcinomas developed. In principle, they can be recorded like the preneoplastic foci of the liver.
  • the detection of an IRS-1 overexpression may thus also for early detection of the renal cancer ⁇ be used.
  • the present invention also relates to a kit for carrying out the methods discussed above, which is characterized in that it contains a reagent specific for IRS-1 as defined above.
  • the kit also contains IRS-1, for example for carrying out a control reaction.
  • the kit also contains other reagents commonly used in immunohistochemical procedures, such as buffers, carriers, markers, etc.
  • Fig. 1 Western blot analysis of liver tissue for the detection of IRS-1 lanes A - D, Western blot analysis for the detection of the 1 65 kD IRS-1 in homogenates of untreated (lane B) and treated with N-nitrosomorpholine (lanes C + D) rat liver tissue. Aliquots (10 ⁇ g) of total cellular protein were subjected to SDS-PAGE under reducing conditions and incubated with polyclonal anti-IRS-1 antibody (Upstate Biotechnology, Lake Placid, NY, USA).
  • the bound antibody was detected with goat anti-rabbit IgG labeled with alkaline phosphatase; Lane A, 3T3 cells (positive control); Lane B, untreated rat liver; Lane C, preneoplastic rat liver; Lane D, hepatocellular carcinoma; left, molecular weight marker
  • glycogen content determined by the PAS reaction, A, E, G- , I, K
  • IRS-1 detected in situ with polyclonal anti-IRS1 antibody (rabbit), B, F, H, J, L
  • APAAP method was confirmed by substitution (C) and control of fluid adsorption (D) in serial frozen sections of preneoplastic (AH) and neoplastic (IL) hepatocellular lesions induced in rats with N-nitrosomorpholine.
  • A preneoplastic glycogenotic foci [ ⁇ ], which show a clearly positive immune response for IRS-1, while the reaction in the surrounding tissue is negative (B, counterstained with cuprolin blue), with substitution (C) and with the controls liquid adsorption, (D);
  • E mixed cell foci [ ⁇ ], which are composed of glycogenotic and low-glycogen cells that show an immune response for IRS-1 (F);
  • G low-glycogen, basophilic cell foci [ ⁇ ] within a glycogenotic focus [ ⁇ ] associated with a positive immune response for IRS-1 in glycogenotic [ ⁇ ] but a negative response in a low-glycogen [ ⁇ ] cell population;
  • I part of a low-glycogen, basophilic hepatocellular carcinoma in the vicinity of MCF [ ⁇ ], associated with a negative immune response for IRS-1 (J), but with a weak immune response in the preneoplastic cell population [ ⁇ ];
  • K part of a hepatocellular carcinoma that contains relatively large amounts of glycogen and
  • PAS periodic acid ship see reaction
  • APAAP alkaline phosphatase anti
  • Example 1 General procedures Animals used and sampling: Preneoplastic hepatocellular lesions were induced in adult male Sprague-Dawley rats by limited exposure to orally administered NNM (stop model; 1 2 mg NNM per kg body weight) over a period of 7 weeks. Tissue from NNM-treated and untreated rats was removed 1 5 and 20 weeks after discontinuation of NNM. HCC was generated by oral administration of the same dose of NNM over a period of 10 weeks or a lower dose (1 mg / kg body weight per day) for up to 73 weeks. Liver tissue was snap frozen at -150 ° C and stored at -80 ° C.
  • Serial frozen sections (6 microns) were stained with Haematox- Ylin and eosin or with SOS-T (periodic acid-Schiff 'see reaction, counterstained with toluidine blue) treated for the detection of glycogen.
  • Example 2 Western blot analysis of liver tissue using anti-IRS-1 antibodies
  • Freeze-dried liver tissue was lysed in buffer containing 0.05 M TBS (Tris-buffered saline), pH 7.4, 1 50 mM NaCl, 1% Nonidet P-40, 1 mM PMSF, 1 mM EGTA, 1 ⁇ g / ml aprotinin, 1 ⁇ g / ml leupeptin, 1 ⁇ g / ml pepstatin, 1 mM Na 3 VO 4 , 1 mM NaF, 2.5% SDS and 5% 2-mercaptoethanol.
  • TBS Tris-buffered saline
  • pH 7.4 Tris-buffered saline
  • PMSF 1 mM PMSF
  • 1 mM EGTA 1 ⁇ g / ml aprotinin
  • 1 ⁇ g / ml leupeptin 1 ⁇ g / ml pepstatin
  • 1 mM Na 3 VO 4 1 mM NaF
  • Protein separation and transfer to a nitrocellulose membrane were carried out using the "PhastSystem” from Pharmacia (Uppsala, Sweden). The membrane was then 20 min. treated with fat-free dry milk (3%) in PBS, 30 min.
  • Frozen sections (6 ⁇ m) were attached to slides coated with poly-L-lysine (Sigma Diagnostics, St. Louis, MO, USA), fixed in 100% methanol (15 min., -20 ° C.), with goat serum, 1 : 30 in TBS, blocked and first 30 min. at RT, then additionally incubated for 24 hours at 4 ° C. with 10 ⁇ g / ml anti-IRS-1 antibody. After each of the steps described below, the frozen sections were run with 0.05 M Tris buffer (pH 7.4) twice for 3 min each. rinsed.
  • the primary antibody was replaced in serial frozen sections with affinity-purified IgG (10 ⁇ g / ml) (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA).
  • affinity-purified IgG 10 ⁇ g / ml
  • a liquid adsorption control was performed in the frozen sections after preincubation of the anti-IRS-1 antibody with the corresponding immunizing peptide (rat IRS-1 peptide, Upstate Biotechnology Inc., Lake Placid, NY, USA) for a period of 3 hours at RT and in a ratio of 1:10 g / g. Both controls turned out to be negative.
  • IRS-1 cannot be detected immunohistochemically in normal liver parenchyma. This result can be explained by the fact that only a very small amount of IRS-1 is contained in normal hepatocytes. In contrast, however, the majority of preneoplastic FAH show a more or less strong immune response to IRS-1. The immune response is limited to the cytoplasm, with a striking perinuclear localization occasionally showing. A total of more than 400 FAH were examined and classified into the classes GSF, MCF and BCF according to the published criteria. The IRS-1 expression was particularly evident in the early GSF (Fig. 2A + B). Of the 93 GSFs examined, 96% were positive for IRS-1.
  • IRS-1 was also clearly detectable by immunohistochemistry (Fig. 2E + F). 97% of the observed MCF were also positive, however the expression of IRS-1 was not quite as strong compared to the GSF.
  • the strength of the Immunreak ⁇ tion with respect to IRS-1 was closely correlated with the number of glycogen storage cells within the FAH.
  • the immune response decreased with the increasing replacement of glycogenotic by low-glycogen basophilic cells, which is an indication that there is a fundamental shift in metabolism during the transition from early to late FAH. This is ⁇ , in conformity with previous observations. FAH made exclusively from basophils Cells passed are hardly observed, but all of the 6 BCFs found were all negative for IRS-1 (Fig. 2G + H).

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Abstract

L'invention concerne des procédés de diagnostic de stades précancéreux du foie et des reins. On détecte la présence du substrat récepteur d'insuline (IRS-1) dans des sites glycogéniques (GSF) et/ou des sites à cellules mélangées (MCF), sur un prélèvement de tissu ou dans les acides nucléiques extraits de ce prélèvement, à l'aide de la liaison d'un réactif spécifique, par exemple, d'un anticorps ou d'une sonde d'acide nucléique, et de la visualisation du complexe formé. La présente invention concerne également un procédé permettant de tester des substances soupçonnées d'induire le cancer du foie ou des reins.
PCT/DE1998/002711 1997-09-09 1998-09-09 Procedes de diagnostic des stades precancereux du foie et des reins WO1999013334A2 (fr)

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DE19739360.8 1997-09-09
DE19739360A DE19739360C2 (de) 1997-09-09 1997-09-09 Verfahren zur Diagnose von frühen Krebsvorstufen der Leber und Niere

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WO1999013334A2 true WO1999013334A2 (fr) 1999-03-18
WO1999013334A3 WO1999013334A3 (fr) 1999-05-27

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002029103A2 (fr) * 2000-10-02 2002-04-11 Gene Logic, Inc. Profils d'expression genetique dans le cancer du foie

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Publication number Priority date Publication date Assignee Title
DE10050338A1 (de) * 2000-10-11 2002-04-25 Deutsches Krebsforsch Auf dem Nachweis von IGF-IRbeta und IRS-1 beruhendes Diagnose- bzw. Klassifizierungsverfahren für Carcinome

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EP0572508A4 (fr) * 1991-01-18 1995-03-29 Joslin Diabetes Center Inc Acide nucleique codant le substrat-1 recepteur d'insuline (irs-1), proteine d'irs-1, maladies et therapie associees au metabolisme de l'irs-1.

Non-Patent Citations (3)

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Title
CHEMICAL ABSTRACTS, vol. 124, no. 13, 25. M{rz 1996 Columbus, Ohio, US; abstract no. 172138, XP002097183 & T. ITO ET AL.: "Overexpression of human insulin receptor substrate 1 induces cellular transformation with activation of mitogen-activated protein kinases." MOLECULAR AND CELLULAR BIOLOGY, Bd. 16, Nr. 3, 1996, Seiten 943-951, Charlestown MA USA *
CHEMICAL ABSTRACTS, vol. 128, no. 10, 9. M{rz 1998 Columbus, Ohio, US; abstract no. 120926, XP002097182 & D. NEHRBASS ET AL.: "Overexpression of insulin receptor substrate-1 emerges early in hepatocarcinogenesis and elicits preneoplastic hepatic glycogenosis." AMERICAN JOURNAL OF PATHOLOGY, Bd. 152, Nr. 2, 1. Februar 1998, Seiten 341-345, Washington DC USA *
MEDLINE, Washington DC USA; abstract no. 97447748, siehe Zusammenfassung XP002097181 & S. TANAKA ET AL.: "Biological effects of human insulin receptor substrate-1 overexpression in hepatocytes" HEPATOLOGY , Bd. 26, Nr. 3, 1. September 1997, Seiten 598-604, Charlestown MA USA *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002029103A2 (fr) * 2000-10-02 2002-04-11 Gene Logic, Inc. Profils d'expression genetique dans le cancer du foie
WO2002029103A3 (fr) * 2000-10-02 2003-09-04 Gene Logic Inc Profils d'expression genetique dans le cancer du foie

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