CN103529220B - CTx is preparing the purposes in Colorectal Cancer Diagnosis medicine - Google Patents
CTx is preparing the purposes in Colorectal Cancer Diagnosis medicine Download PDFInfo
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- 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
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- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
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- G01N2800/56—Staging of a disease; Further complications associated with the disease
Abstract
The present invention relates to biological technical field, the c-terminal peptides CTx particularly relating to a kind of extracellular matrix components type i collagen is preparing the purposes in Colorectal Cancer Diagnosis medicine.The invention provides CTx and prepare the purposes in Colorectal Cancer Diagnosis medicine, Screening Diagnosis colorectal cancer medicine and screening treatment colorectal cancer medicine.The present invention chooses Colorectal Carcinoma, serum clinical samples, and research type i collagen and CTx, in colorectal cancer, development and the clinical value by stages in prognosis etc. occur.Pass through great many of experiments, inventor finds that CTx is in difference by stages Patients with Colorectal Cancer serum, change and the pathological staging of expression present positive correlation, and CTx is compared with existing tumor in digestive tract carcinomebryonic antigen, better can distinguish transfer and non-diverting Patients with Colorectal Cancer.In addition, inventor also finds that CTx and Patients with Colorectal Cancer 3 years disease free survivals are relevant, and its high expressed imply that Patients with Colorectal Cancer 3 years is bad without knurl survival region.
Description
Technical field
The present invention relates to biological technical field, the c-terminal peptides CTx particularly relating to a kind of extracellular matrix components type i collagen is preparing the purposes in Colorectal Cancer Diagnosis medicine.
Background technology
Collagen is as extracellular matrix (Extracellular matrix, ECM) one of main composition, in most of connective tissue, the structural stability maintaining ECM is played an important role, to many fiber diseases as relevant (Lai such as liver fibrosis, K.K., Shang, S., Lohia, N., etc.Extracellular matrix dynamics in hepatocarcinogenesis:acomparative proteomics study of PDGFC transgenic and Pten null mouse models.PLoS Genet., 2011, 7 (6): e1002147.).Recent report also points out invasion inhibition also extremely important (Sund, M., the Kalluri of collagen for tumour cell, R.Tumor stroma derived biomarkers in cancer.Cancer Metastasis Rev., Jun, 2009,28 (1-2): 177-83.).Be reported in human body at present and had 28 kinds of collagens and exist or may exist, and wherein type i collagen is that people's in-vivo content distributes at most the widest a kind of collagen.
Type i collagen is the triple helix structure formed by two α 1 chains and α 2 chain, is generally synthesized by fibroblast, as a part for tumor microenvironment, plays a significant role in tumor cell proliferation, transfer process.Find in colorectal cancer research, type i collagen high expressed is in presenting (Oku Y in the tumour cell infiltrating growth, Shimoji T, Takifuji K, etc.Identification of the molecular mechanisms for dedifferentiation at the invasion front of colorectalcancer by a gene expression analysis.Clin Cancer Res, 2008,14:7215 – 7222.).Can reduce E-cadherin and β-catenin expression in vitro in clone makes cell and intercellular adhesion decline, cell is impelled to occur stem-like cell phenotype (SCKirkland.Type I collagen inhibits differentiation and promotes a stem cell-like phenotype in humancolorectal carcinoma cells.Br J Cancer.2009, 101 (2): 320-6.), and Cdx2 can be reduced express thus T suppression cell differentiation (Brabletz, T., Spaderna, S., Kolb, J., etc.Down-regulation of the homeodomain factorCdx2in colorectal cancer by collagen type I:an active role for the tumor environment in malignanttumor progression.Cancer Res., 2004, 64 (19): 6973-7.).In addition, type i collagen is by promoting malignant transformation (the Grzesiak J of pancreatic cancer cell with a2 β 1integrin effect, Bouvet M.The a2 β 1integrin mediates themalignant phenotype on type I collagen in pancreatic cancer cell lines.Br J Cancer, 2006, 94:1311 – 1319), also be one of 17 and the closely-related gene of metastases (Ramaswamy S simultaneously, Ross KN, LanderES, etc.A molecular signature of metastasis in primary solid tumors.Nat Genet.2003, 33 (1): 49-54.).Although above-mentioned in cell model the effect of type i collagen to tumour be widely studied, at present less from the expression study in malignant tumor tissue for type i collagen.
When there is matrix reconstruction (ECM remodeling), type i collagen can be degraded, and its N holds and the end peptide (N-telopeptides and C-telopeptides) of C end can be discharged in blood.CTx is exactly type i collagen C end peptide the preceding paragraph 8 amino acid whose fragments (EKAHDGGR), can be undertaken detecting (Bonde by ELISA in serum as the index of type i collagen degraded, M., Garnero, P., Fledelius, C., etc.Measurement of bone degradation products inserum using antibodies reactive with an isomerized form of an8amino acid sequence of theC-telopeptide of type I collagen.J Bone Miner Res, 1997, 12 (7): 1028-34.).Research for CTx is studied more in osteopathology, and CTx is also at present as judging the reasonable biomarker (Rosen of bone information (Bone resorption), H.N., Moses, A.C., Garber, J., etc.Serum CTX:a new marker of boneresorption that shows treatment effect more often than other markers because of low coefficient ofvariability and large changes with bisphosphonate therapy.Calcif Tissue Int, 2000, 66 (2): 100-3.Marx, R.E., Cillo, J.E., Jr., Ulloa, J.J.Oral bisphosphonate-induced osteonecrosis:risk factors, prediction of risk using serum CTX testing, prevention, and treatment.J Oral Maxillofac Surg, 2007, 65 (12): 2397-410.).At present about in the report of itself and cancer-related, in blood, the rise of CTx is at breast cancer (Cloos, P.A., Christgau, S., Lyubimova, N., etc.Breast cancer patients with bone metastases arecharacterised by increased levels of nonisomerised type I collagen fragments.Breast Cancer Res, 2003, 5 (4): R103-9.), prostate cancer (Garnero, P., Buchs, N., Zekri, J., etc.Markers of bone turnover forthe management of patients with bone metastases from prostate cancer.Br J Cancer, 2000, 82 (4): 858-64.) and lung cancer (Leeming, D.J., Koizumi, M., Byrjalsen, I., etc.The relative use of eightcollagenous and noncollagenous markers for diagnosis of skeletal metastases in breast, prostate, orlung cancer patients.Cancer Epidemiol Biomarkers Prev, 2006, 15 (1): 32-8.) report is had in Bone tumour patient.But except as the standard judging bone metastaes, the clinical value of CTx in other tumor development process and in diagnosis of colorectal carcinoma prognosis etc. is not appeared in the newspapers.
Summary of the invention
The shortcoming of prior art in view of the above, the object of the present invention is to provide the purposes of a kind of c-terminal peptides CTx of extracellular matrix components type i collagen in preparation or Screening Diagnosis colorectal cancer medicine, for solving the problems of the prior art.
For achieving the above object and other relevant objects, first aspect present invention provides CTx preparing the purposes in Colorectal Cancer Diagnosis medicine, Screening Diagnosis colorectal cancer medicine and screening treatment colorectal cancer medicine.
Described Screening Diagnosis colorectal cancer medicine specifically refers to use CTx, to the assessment may carrying out biologically active, pharmacological action and medical value as the material of Colorectal Cancer Diagnosis drug use.
Described screening treatment colorectal cancer medicine specifically refers to use CTx, to the assessment may carrying out biologically active, pharmacological action and medical value as the material for the treatment of colorectal cancer drug use.
CTx in the present invention is 8 amino acid peptide sections of type i collagen α 1 chain carboxy-terminal, and its amino acid sequence is EKAHDGGR(Seq ID No.1).When being in cell generation matrix reconstruction under some physiology or pathological state, type i collagen in extracellular matrix can be degraded, its N holds and the end peptide of C end can be discharged in blood, and described CTx holds one of fragments of peptides for the type i collagen C end discharged when type i collagen degrade.
Preferably, described Colorectal Cancer Diagnosis specifically refers to colorectal cancer preoperative diagnosis by stages or to colorectal cancer Index for diagnosis.(mainly contain help not need before surgery can to proactive judgement by stages by pathology detection)
According to and combine the TNM(tumor-node-metastasis of formulation according to International Union Against Cancer (UICC) and tumour federation of the U.S. (AJCC)) allotment method, colorectal cancer can be divided into I-IV phase four-stage, and described staging of colorectal cancer specifically refers to which stage distinguishes colorectal cancer is according to described TNM allotment method.
Describedly specifically refer to carry out Index for diagnosis, more specifically for carry out Index for diagnosis to the disease free survival of Patients with Colorectal Cancer to the course of disease of Patients with Colorectal Cancer and final result to colorectal cancer Index for diagnosis.
Preferably, described colorectal cancer is Human colorectal carcinoma.
Preferably, described CTx is biomarker.
Preferred, described CTx is serum biomarkers.
Preferably, the type i collagen C end fragments of peptides of described CTx for discharging during type i collagen degraded during matrix reconstruction.
Preferred, described type i collagen derives from people.
Because in clinical diagnostics, I, II phase belongs to the commitment of colorectal cancer, if now find, tumour can be cured by excision mostly, III phase patient has started to find lymphatic metastasis, there is far-end transfer in IV phase patient, four period five-year survival rate reduce successively, recurrence rate after healing increases progressively.Because CTx just can be used as early stage specific biological mark, its discovery is significant for the prediction prognosis of colorectal cancer.
Second aspect present invention provides a kind of Colorectal Cancer Diagnosis biomarker, and described biomarker is CTx, and the amino acid sequence of described CTx is as shown in Seq ID No.1.
Preferably, described biomarker is serum biomarkers.
Preferably, described Colorectal Cancer Diagnosis specifically refers to colorectal cancer preoperative diagnosis by stages or to colorectal cancer Index for diagnosis.
Preferably, described colorectal cancer is Human colorectal carcinoma, the type i collagen C end fragments of peptides of described biomarker CTx for discharging when type i collagen during matrix reconstruction (people source) is degraded.
The present invention chooses Colorectal Carcinoma, serum clinical samples, and research type i collagen and CTx, in colorectal cancer, development and the clinical value by stages in prognosis etc. occur.Pass through great many of experiments, inventor finds that CTx is in difference by stages Patients with Colorectal Cancer serum, change and the pathological staging of expression present positive correlation, and CTx and existing tumor in digestive tract carcinomebryonic antigen (Carcinoembryonic antigen, CEA) compare, better can distinguish transfer and non-diverting Patients with Colorectal Cancer.In addition, inventor also finds that CTx and Patients with Colorectal Cancer 3 years disease free survivals are relevant, and its high expressed imply that Patients with Colorectal Cancer 3 years is bad without knurl survival region.
In addition, in the present invention, inventor further provides the protein I Collagen Type VI of this end peptide of generation CTx in Expression in Colorectal Cancer situation, reference frame is provided for understanding the effect of extracellular matrix in colorectal cancer generation evolution in depth, and find thus, the change and the pathological staging that generate the expression of the protein I Collagen Type VI of this end peptide of CTx there is no obvious correlativity, further demonstrate that CTx is preparing the using value in Colorectal Cancer Diagnosis medicine thus.
Accompanying drawing explanation
Fig. 1 represents the expression of type i collagen in difference Colorectal Carcinoma mRNA level in-site by stages.
Fig. 2 represents the expression of type i collagen at difference Colorectal Carcinoma protein level by stages.
Fig. 3 represents that type i collagen is in difference Colorectal Carcinoma immunohistochemical staining result by stages.
Fig. 4 represents that type i collagen degraded index CTx and random degenerate small fragment COL1 ELISA in normal and difference by stages Patients with Colorectal Cancer serum detects.
Fig. 5 represents that CTx and CEA compares can better distinguish be shifted and non-diverting Patients with Colorectal Cancer.
Fig. 6 represents that CTx and Patients with Colorectal Cancer 3 years disease free survivals are relevant, and its high expressed imply that Patients with Colorectal Cancer 3 years is bad without knurl survival region.
Fig. 7 represents that CTx and Patients with Colorectal Cancer 3 years disease free survivals are relevant, and its expression and Patients with Colorectal Cancer 3 years disease free survivals are negative correlativing relation.
Embodiment
Below by way of specific instantiation, embodiments of the present invention are described, those skilled in the art the content disclosed by this instructions can understand other advantages of the present invention and effect easily.The present invention can also be implemented or be applied by embodiments different in addition, and the every details in this instructions also can based on different viewpoints and application, carries out various modification or change not deviating under spirit of the present invention.
Before further describing the specific embodiment of the invention, should be understood that protection scope of the present invention is not limited to following specific specific embodiments; It is also understood that the term used in the embodiment of the present invention is to describe specific specific embodiments, instead of in order to limit the scope of the invention; In instructions of the present invention and claims, unless explicitly pointed out in addition in literary composition, singulative " ", " one " and " this " comprise plural form.
When embodiment provides numerical range, should be understood that except non-invention is otherwise noted, between two end points of each numerical range and two end points, any one numerical value all can be selected.Unless otherwise defined, all technology used in the present invention are identical with the meaning that those skilled in the art of the present technique understand usually with scientific terminology.Except the concrete grammar used in embodiment, equipment, material, according to those skilled in the art to the grasp of prior art and record of the present invention, any method of prior art that is similar with the method described in the embodiment of the present invention, equipment, material or that be equal to, equipment and material can also be used to realize the present invention.
Unless otherwise indicated, disclosed in the present invention experimental technique, detection method, preparation method all adopt the routine techniques of the molecular biology of the art routine, biological chemistry, chromatin Structure and analysis, analytical chemistry, cell chulture, recombinant DNA technology and association area.These technology are existing in existing document improves explanation, specifically can see the MOLECULAR CLONING:A LABORATORY MANUAL such as Sambrook, Second edition, Cold Spring HarborLaboratory Press, 1989and Third edition, 2001; Ausubel etc., CURRENT PROTOCOLS INMOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987and periodic updates; The seriesMETHODS IN ENZYMOLOGY, Academic Press, San Diego; Wolffe, CHROMATINSTRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; METHODSIN ENZYMOLOGY, Vol.304, Chromatin (P.M.Wassarman and A.P.Wolffe, eds.), AcademicPress, San Diego, 1999; With METHODS IN MOLECULAR BIOLOGY, Vol.119, ChromatinProtocols (P.B.Becker, ed.) Humana Press, Totowa, 1999 etc.
Inventor found in colorectal cancer proteomic efforts in early stage, and type i collagen is high expressed in difference by stages Patients with Colorectal Cancer serum, showed that type i collagen may be closely related with colorectal cancer.Inventor has found the high expressed of type i collagen in difference by stages Colorectal Carcinoma, and in serum, detect the expression of type i collagen different fragments in difference by stages Patients with Colorectal Cancer further, find its c-terminus degraded end PEPC Tx and Patients with Colorectal Cancer by stages and prognosis relevant, complete the present invention on this basis.One, type i collagen detection in the tissue
Embodiment 1: the collection of different Patients with Colorectal Cancer sample by stages and normal sample
Year May in May, 2009 to 2010, we collect the tissue of 91 routine Patients with Colorectal Cancers and serum sample altogether, and (patient has been clarified a diagnosis by histopathology, and it suffers from colorectal cancer, and the TNM(tumor-node-metastasis of formulation is combined according to International Union Against Cancer (UICC) and tumour federation of the U.S. (AJCC)) allotment method determines pathological staging, wherein I phase 21 people, II phase 41 people, III phase 22 people, IV phase 7 people, all patients have all carried out laparoscopic surgical procedure), have collected the healthy human body serum sample of 33 examples without disease of digestive tract and diseases associated with inflammation simultaneously.Within every 3 months, carry out tracking in all patients after surgery first 2 years to pay a return visit, within postoperative 3rd year every 6 months, carry out trackings and pay a return visit, stopped to postoperative 41st month, record recurrence or the death time point of patient.Patients clinical information is as shown in table 1.
Table 1 patients clinical information
aaC, the colon ascendens; TC, transverse colon; DC, colon descendens; SC, sigmoid colon; RC, rectum;
Embodiment 2:I Collagen Type VI is in the expression of difference Colorectal Carcinoma RNA and protein level by stages
1, RNA is organized to extract and real-time fluorescence quantitative PCR (Realtime PCR) detection:
We are extracted total mRNA of tumor tissues and corresponding normal colorectal carcinoma, and have detected the mrna expression level of its type i collagen respectively.Total mRNA of tissue is extracted with the Trizol kit of Invitrogen company, use the PrimeScript RT reagent Kit kit of Takara company from the RNA reverse transcription synthesis 20ml cDNA of 2mg, 7500 real-time fluorescence quantitative PCR instrument of AppliedBiosystems company and corresponding SYBR Green PCR Master Mix kit is used to carry out real-time fluorescence quantitative PCR (adopt 2-△ △ Ct analytic approach to analyze gene expression abundance, GAPDH is as reference gene).The primer sequence that PCR reaction uses is as shown in table 2.(result is as Suo Shi Fig. 1 and table 3)
The primer sequence that table 2PCR reaction uses
2, histone extracts and Western blotting (Western Blot) detection
For carrying out the protein expression analysis of tumor tissues and normal colorectal carcinoma, freezing tissue samples 100mg is added RIPA lysate (50mM Tris-HCl (pH7.4), 0.4wt%sodium deoxycholate, 150mM NaCl, 1wt%Triton, 1mM PMSF) after to carry out the broken and cracking of about 30 minutes homogenates on ice, afterwards at 4 DEG C, 16, under 000 × g condition centrifugal 30 minutes, carry out the protein quantification of solution by BCA method.Use β-actin as the quantitative internal reference of loading; the equal protein (50mg/ band) of each sample is carried out 8wt%SDS-PAGE electrophoretic separation; be transferred on the nitrocellulose filter of GE company afterwards; primary antibodie 4 DEG C of overnight incubation (anti-collagen I (1:5000, ab34710 is used after closing; Abcam).Immune response positive band uses Li-COR company Odyssey Infrared Imaging System to carry out observing and taking pictures, and uses Quantity One to carry out quantitatively.(result is as Fig. 2)
3, histogenic immunity groupization detects
The tissue paraffin section de of 3 – 5mm uses 4wt% pepsin in 37 DEG C of reaction 20min, and carry out antigen retrieval, spend the night with primary antibodie 4 DEG C of reactions (anti-collagen I, 1:500, ab34710 afterwards; Abcam), the Envisiondetection system of DAKO company is used to carry out observing and taking pictures.(result is as Fig. 3)
4, interpretation of result
Table 3I Collagen Type VI is in the real-time fluorescence quantitative PCR testing result raw data of difference Colorectal Carcinoma by stages
| Sample number into spectrum | Pathological staging | COL1A1(tumour/normal) | COL1A2(tumour/normal) |
| 88 | I | 17.09 | 5.99 |
| 51 | I | 3.04 | 1.81 |
| 74 | I | 3.01 | 1.57 |
| 101 | I | 1.81 | 1.12 |
| 72 | I | 1.69 | 0.91 |
| 116 | I | 266.35 | 30.92 |
| 67 | II | 124.16 | 33.06 |
| 48 | II | 113.29 | 32.96 |
| 79 | II | 16.42 | 10.02 |
| 59 | II | 15.11 | 7.22 |
| 84 | II | 11.31 | 7.03 |
| 75 | II | 7.61 | 4.91 |
| 47 | II | 7.11 | 1.04 |
| 55 | III | 18.33 | 14.49 |
| 17 | III | 4.81 | 3.22 |
| 68 | III | 4.36 | 2.93 |
| 82 | III | 3.18 | 2.00 |
| 62 | III | 1.91 | 1.86 |
| 39 | III | 1.34 | 1.74 |
| 81 | III | 1.14 | 0.98 |
| 94 | IV | 3.43 | 3.34 |
| 61 | IV | 1.21 | 1.58 |
| 53 | IV | 1.27 | 1.04 |
| 103 | IV | 0.94 | 0.59 |
| 44 | IV | 1.41 | 0.44 |
Visible from the result of table 3 and Fig. 1, Fig. 2, no matter at mRNA level in-site or protein level, in each phase colon cancer tumours tissue, type i collagen is expressed all high than normal structure, illustrates that type i collagen is up-regulated expression really in tumor tissues when cancer occurs.But, can find out from the result of Fig. 1 and Fig. 2, type i collagen the non-diverting phase (I phase and II phase) than transfer phase (III phase and IV phase) tumor tissues in high expressed, especially on protein level, there is significant difference (p<0.05).
Type i collagen expression is expressed the highest in II phase tumor tissues, and on protein level, its tumor tissues of COL1A1 and COL1A2 and normal structure ratio reach 22.54 and 7.19 times respectively.In clinical diagnostics, I, II phase belongs to commitment, if now find, tumour can be cured by excision mostly, III phase patient has started to find lymphatic metastasis, and far-end transfer occurs IV phase patient, four period five-year survival rate reduce successively, recurrence rate after healing increases progressively.Therefore, at clinical middle commitment, especially the II phase is critical stage in the evolution of colorectal cancer, the finding that there is and help predict prognosis, in important significance in clinical application of clinical stages diagnostic biomarkers.In addition, the albumen of some II phase high expresseds probably moves relevant with the aggressive of tumour cell.
Two, the detection of CTx in serum
Embodiment 3:I Collagen Type VI degraded index CTx and random degenerate small fragment COL1 ELISA in normal and difference by stages Patients with Colorectal Cancer serum detects:
1, ELISA detection kit and method
COL1 and CTx level in serum can use commercial reagents box (Uscn Life Science & Technology Co., Houston, TX, USA) ELISA detection is carried out, change of serum C Tx uses the being at war with property of specific monoclonal antibody of linear EKAHDGGR peptide section (fragment of the α 1 chain carboxy-terminal peptide of type i collagen) to suppress ELISA to detect, in brief, in biotin labeled exactly synthesis CTx peptide section and serum, unlabelled CTx is suppressed to react by good special being at war with property of CTx antibody with wrapping in advance, horseradish peroxidase Avidin is added in each reaction micropore, hatch, concrete detection method reference reagent box instructions.After adding substrate solution, in the color of reaction mixture and serum, the concentration of CTx is negative correlation.In serum, CEA level uses commercial reagents box to carry out detecting (Roche Diagnostics, Mannheim, Germany), concrete detection method reference reagent box instructions.
2, Analysis of test results
CTx is respectively 2.19,3.16 from the I phase to the average serum content of IV phase, and 3.58 and 4.58ng/ml, the change of its expression and the pathological staging of colorectal cancer present positive correlation.In terminal cancer (III+IV phase), the level of CTx is significantly higher than the respective horizontal (p=0.003, Mann – Whitney U test) of CTx in early-stage cancer (I+II phase).(result is as Fig. 4)
3, CTx and staging of colorectal cancer and Analysis of relationship between prognosis
Good positive correlation is there is between the pathological staging due to CTx and colorectal cancer, and it is by stages all closely related with the early diagnosis of cancer patient, result for the treatment of and prognosis information, therefore we so that receiver operator curve (Receiveroperating characteristic has been carried out to CTx, and 3 years disease free survival analysis (Disease-free survival ROC), DFS), assessment is made in order to the clinical value potential to CTx.Result can be found out, CTx is better than the differentiation effect (AUC:0.584) of CEA for differentiation effect (AUC:0.696) that is early stage, end-stage patients.(result is as Fig. 5)
In addition we have evaluated CTx and colorectal cancer survive without knurl between contact, with the mean value 4.08ng/ml of all patient CTx for boundary, patient is divided into two groups.Kaplan-Meier survival analysis result shows, high expressed group 3 years disease free survivals of CTx are starkly lower than another group (log rank test, p=0.006), CTx is carried out the analysis of Cox proportional hazard model and also illustrate that CTx can provide important information for the Index for diagnosis of Patients with Colorectal Cancer.(result is as Fig. 6)
Further, experimenter is divided into 3 groups by the serum content of CTx, is respectively 0<CTx≤2,2<CTx≤4,4<CTx; (log rank test, p=0.022), Kaplan-Meier survival analysis result shows, along with CTx expresses the lifting gradually of concentration, 3 years disease free survivals of Patients with Colorectal Cancer reduce gradually, both present negative correlativing relation, further demonstrate that CTx can be the important value of the Index for diagnosis of Patients with Colorectal Cancer.(result as shown in Figure 7)
In sum, the present invention effectively overcomes various shortcoming of the prior art and tool high industrial utilization.
Above-described embodiment is illustrative principle of the present invention and effect thereof only, but not for limiting the present invention.Any person skilled in the art scholar all without prejudice under spirit of the present invention and category, can modify above-described embodiment or changes.Therefore, such as have in art usually know the knowledgeable do not depart from complete under disclosed spirit and technological thought all equivalence modify or change, must be contained by claim of the present invention.
Claims (5)
1.CTx is preparing purposes in Screening Diagnosis colorectal cancer medicine of the purposes in Colorectal Cancer Diagnosis medicine, CTx or the purposes of CTx in screening treatment colorectal cancer medicine, described CTx is serum biomarkers, and the purposes in described Screening Diagnosis colorectal cancer medicine and the screening purposes for the treatment of in colorectal cancer medicine do not comprise diagnosis or the methods for the treatment of of disease.
2. purposes as claimed in claim 1, is characterized in that, described Colorectal Cancer Diagnosis specifically refers to colorectal cancer preoperative diagnosis by stages or to colorectal cancer Index for diagnosis.
3. purposes as claimed in claim 1, it is characterized in that, described colorectal cancer is Human colorectal carcinoma.
4. purposes as claimed in claim 1, is characterized in that, the type i collagen C end fragments of peptides of described CTx for discharging when type i collagen is degraded.
5. purposes as claimed in claim 4, is characterized in that, the type i collagen C end fragments of peptides of described CTx for discharging during type i collagen degraded during matrix reconstruction.
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