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Human colorectal carcinoma molecular marker COL3A1 and application thereof

A technology of colon cancer and markers, applied in the fields of genetic engineering and medical detection, which can solve problems such as interference

Inactive Publication Date: 2017-02-15
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Secondly, conventional methods rely on pathological parameters, TNM staging, Dukes staging, and other methods for predicting the prognosis and survival of patients after surgery or radiotherapy and chemotherapy. However, human factors often interfere with pathological judgments. Molecular markers with significant differences, high sensitivity, good specificity, and easy operation are used as auxiliary judgment methods. Therefore, it is an urgent task for medical researchers to find effective markers for colon cancer and accurately judge the clinical stage and prognosis of colon cancer.

Method used

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  • Human colorectal carcinoma molecular marker COL3A1 and application thereof
  • Human colorectal carcinoma molecular marker COL3A1 and application thereof
  • Human colorectal carcinoma molecular marker COL3A1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Using the Oncomine public database ( https: / / www.oncomine.org / resource / login.html ) to analyze the expression of COL3A1 gene in colon cancer and para-cancerous tissues, the analysis method: log on to the above-mentioned Oncomine database, input the SPON2 gene name, select colon cancer as the tumor type, select mRNA as the data type, select cancer and normal analysis as the differential analysis type, and then analyze For the expression of COL3A1 gene in each data set, download the expression values ​​of COL3A1 gene in cancer and paracancerous tissues in each data set, and analyze whether the difference in expression is significant; Shows a significant high expression trend, which is evidenced by the analysis of multiple data sets, including GaedckeColorectal ( figure 1 -A, n=65, p=8.5E-13), Hong Colorectal (Figure-1B, n=70, p=0.037), Kaiser Colon (Figure-1C, n=100, p=0.029), Skrzypczak Colorectal 2 (Figure-1D, n=10, p=0.003 for the ratio of cancer to paracancer) and T...

Embodiment 2

[0046] Example 2 Western blot method was used to detect the expression of COL3A1 protein in colon cancer cell lines,

[0047] 1. Main solution preparation:

[0048] 1) PBS solution: Measure 25mL of Sangon 20×PBS solution, dilute to 500mL with ultrapure water, store at room temperature,

[0049] 2) 50×Protease inhibitor cocktail (protease inhibitor mother solution): take a Roche protease inhibitor (complete EDTA-free Protease Inhibitor cocktail tablets), add 1mL ultrapure water to dissolve, subpackage, store at -20°C, this is 50× protease inhibitor stock solution,

[0050] 3) Cell lysate: 50mM Tris-HCl (pH7.4), 1% SDS, 2mM EDTA, aliquoted, stored at -20°C, add 20μL 50×Protein inhibitor cocktail for each ml of lysate,

[0051] 4) Acrylamide solution: 30% acrylamide, 0.8% methylene bisacrylamide,

[0052] 5) Separating gel buffer: 1.5mol / L Tris-HCl pH 8.8,

[0053] 6) Stacking gel buffer: 1.0mol / L Tris-HCl pH 6.8,

[0054] 7) SDS: 10% (m / v),

[0055] 8) Ammonium persulfate ...

Embodiment 3

[0094] Example 3 Analysis of the expression of COL3A1 protein in colon cancer and adjacent normal tissues by immunohistochemistry

[0095] 1. Tissue chip: Using a commercial tissue chip (Shanghai Xinchao Biotechnology Co., Ltd.), HCol-Ade180Sur-04 includes 90 cases, 180 points of cancer and paracancerous tissues, and clinical information includes gender, age, TNM, tumor size, stage, and grade , follow-up information, etc. The diameter of the chip sample point is 1.5mm, and the fixing method is formalin;

[0096] 2. Baking wax

[0097] 1) Put the tissue chip into the oven, adjust the temperature to 63 degrees, and bake the wax for one hour;

[0098] 2) Reagent preparation: 10×PBS buffer solution (recipe: 80g NaCl, 2g KCl, 15.35g Na 2 HPO 4 , 2gKH 2 PO 4 , dilute to 1000 ml with pure water): Dilute 10×PBS buffer to 1×PBS buffer, then add 0.05% Tween reagent to the total volume in 1×PBS buffer; antigen retrieval solution: 82mL 0.1 mol / L sodium citrate solution + 18mL 0.1mo...

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Abstract

The invention belongs to the fields of genetic engineering and medical detection, and relates to a human colorectal carcinoma molecular marker COL3A1 and an application thereof in preparing a human colorectal carcinoma detecting preparation or a detecting kit. On the basis of an experiment of analyzing the expression of a COL3A1 gene in colorectal carcinoma and para-carcinoma tissues, a result shows that the COL3A1 gene has a more significant high-expression trend in the carcinoma tissues in comparison with that in the para-carcinoma tissues, proving that the COL3A1 plays a role in the occurrence and development of the colorectal carcinoma; the COL3A1 presents significant different expression in colorectal carcinoma and para-carcinoma epithelial cells but has no expression difference in carcinoma and para-carcinoma matrix parts, showing that the COL3A1 protein is the carcinoma epithelial cell specificity molecular marker; and meanwhile, up-regulation of the expression of the COL3A1 gene is positively related to tumor progression and is related to the postoperative survival time of a patient with the colorectal carcinoma. The invention provides a scientific basis for the effective judgment of canceration progression of the human colorectal carcinoma.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and medical detection, and relates to a human colorectal cancer molecular marker COL3A1 and its use, in particular to its use in preparing human colorectal cancer detection preparations or detection kits. Background technique [0002] According to the World Health Organization GLOBOCAN2012 statistical report, colon cancer has become the third largest male tumor and the second largest female tumor worldwide, and its prognosis and survival are very poor. Among them, the overall mortality rate in 2012 was 50.1% (male) and 52.1% (women). Tumor occurs and affects individual families, and brings serious social impact, increasing public health expenditure and financial burden. Therefore, early detection, early confirmation, and early treatment are of great significance for improving the cure rate of colon cancer and prolonging the prognosis and survival time. Studies have shown that the initial stag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574
Inventor 刘锋杨芃原王晓庆崔书建姜英华
Owner FUDAN UNIV
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