Detection method of marker for diagnosing colorectal cancer

A detection method and technology of colorectal cancer, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as the unsystematic establishment of detection methods

Pending Publication Date: 2021-06-29
广州医科大学附属中医医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the detection method of DCD in colorectal can

Method used

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  • Detection method of marker for diagnosing colorectal cancer
  • Detection method of marker for diagnosing colorectal cancer
  • Detection method of marker for diagnosing colorectal cancer

Examples

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Embodiment 1

[0031] A detection method for a marker for diagnosing colorectal cancer, comprising the following steps:

[0032] 1) Select 5 groups of colorectal cancer tissues (T), select normal human cells next to the 5 groups of colorectal cancer tissues, that is, paracancerous cells (N), that is, select 5 pairs of colorectal cancer tissues (T) and paracancerous tissues (N) As a sample, the extraction method of total protein is as follows: After cutting 0.1g tissue sample, put it into a pre-cooled homogenizer, add RIPA lysate containing 1.5% PMSF at a ratio of 1:10, and heat it at 4-10°C Homogenize until the tissue is eliminated, transfer the sample to a 1.5nL centrifuge tube, centrifuge at a temperature of 4-10°C and a centrifugal force of 12,000g for 10 minutes, absorb the supernatant, and obtain the total protein of the tissue sample; the storage method of the total protein is as follows: Add 5×Loading buffer to the supernatant, heat at 80-100°C for 5-10 minutes, and store the total pr...

Embodiment 2

[0043] A detection method for a marker for diagnosing colorectal cancer, comprising the following steps:

[0044] 1) Select five intestinal cancer cell lines and one human normal intestinal epithelial cell (HIEC) as cell samples, among which five intestinal cancer cell lines are SW620 cells, LoVo cells, HT-29 cells, HCT-116 cells and SW480 cells, The method for extracting the total protein of the cells is as follows: wash away the cell culture medium of a certain cell sample containing serum with PBS respectively, and after the liquid is sucked dry, press 1×10 6 Add 200 μl of RIPA lysate containing 1.5% PMSF at a cell density of 1 cell / ml, and blow with a pipette at a temperature of 4°C to fully lyse the cells to obtain total protein. Centrifuge at a temperature of 4°C and a centrifugal force of 12,000g for 10 minutes. , absorb the supernatant; the storage method of the total protein is: add 5×Loading buffer to the supernatant, heat at 100°C for 5min, and store at -20°C;

[0...

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Abstract

The invention relates to the technical field of in-vitro detection, in particular to a detection method of a marker for diagnosing colorectal cancer. The detection method comprises the following steps of (1) extracting the total protein of cells or tissues; (2) preparing a separation gel and a concentrated gel; (3) performing polyacrylamide gel electrophoresis; (4) transferring a film; (5) incubating an antibody; and (5) developing by a chemiluminescence method. According to the present invention, the content of DCD in tissues or cells is detected through western-blot, so that whether the tissues or cells are intestinal cancer tissues or intestinal cancer cells or not is judged, DCD is a candidate diagnostic index and therapeutic target of colorectal cancer, and a new diagnostic thought can be provided for clinic.

Description

technical field [0001] The invention relates to the technical field of in vitro detection, in particular to a detection method for a marker for diagnosing colorectal cancer. Background technique [0002] DCD (Dermcidin) is a natural active peptide isolated from human sweat by German scientists Schittek et al. in 2001. DCD was initially thought to be specifically expressed in sweat glands, expressed in sweat glands and secreted into sweat. antibacterial activity. Later studies found that DCD is also expressed in certain cancers, including melanoma, skin tumors, breast, prostate, pancreas, lung and hepatocellular carcinoma. DCD is likely to be a potential oncogenic factor and could serve as a candidate therapeutic target for the treatment of these cancers. [0003] Experimental studies have confirmed that DCD is highly expressed in colorectal cancer, which ranks third in the incidence of tumors in the world, and can become a diagnostic index and therapeutic target for colore...

Claims

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Application Information

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IPC IPC(8): G01N33/574
CPCG01N33/57446
Inventor 邱芳华李秋明刘道利龙华婧
Owner 广州医科大学附属中医医院
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