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Method for detecting proteome in rectal cancer

A technology of proteomics and colorectal cancer, applied in the field of medical biology, can solve the problems of reduced reliability of functional research results, and achieve the effects of avoiding heterogeneity, improving reliability, and wide detection range

Active Publication Date: 2018-03-30
JINAN UNIVERSITY
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Problems solved by technology

Lead to a decrease in the reliability of subsequent functional research results, etc.

Method used

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  • Method for detecting proteome in rectal cancer
  • Method for detecting proteome in rectal cancer
  • Method for detecting proteome in rectal cancer

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Embodiment 1

[0057] In this case, human colorectal cancer cells HCT116 and RKO were used to screen I8 cells with high invasive ability by Transwell method combined with SILAC quantitative proteomics technology and IPA bioinformatics analysis to find the effect of Cdc42 signaling pathway and protein Cdc42BPA on the invasion and metastasis of colorectal cancer.

[0058] Transwell Screening Cells

[0059] (1) Rinse wild-type HCT116 and RKO cells twice with 1×PBS, digest and collect them with 0.05% trypsin, centrifuge to remove the supernatant, and resuspend the cells with serum-free medium (1640);

[0060] (2) Cell counting, according to the formula: cell density (unit / mL) = (total number of 4 cells / 4) × 10 4 ×Dilution factor;

[0061] (3) Take 20×10 4 Add cells to the upper chamber, and fill the volume difference between groups with serum-free DMEM medium;

[0062] (4) Add 600 μL of culture solution containing 20% ​​serum under the small chamber;

[0063] (5) Placed at 37°C, 5% CO 2 Inc...

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Abstract

The invention relates to research on cancer cell metastasis, and particularly discloses a method for detecting a proteome in rectal cancer. According to the method, highly invasive rectal cancer cellsHCT116-I8 are screened by utilizing Transwell, and a protein or a pathway relating to cell invasion capacity is found by performing quantitative protein identification by combining an SILAC technology. Dynamic expression level of the protein in cells with high invasion capacity can be analyzed by a proteomic technology from an overall perspective, and a protein expression profile can be detectedpanoramically. The changes of proteomes in the cells with different invasion capacity in the rectal cancer can be detected in a high-throughput, high-precision and high-resolution manner by utilizinga highly invasive intestinal cancer model and a technology of screening invasion related functional protein by proteomics, and a protein pathway relating to invasion and metastasis can be found. The occurrence mechanism of diseases, such as cancer, can be explored efficiently by utilizing the technology, and a medicament acting target with high reliability is provided.

Description

technical field [0001] The invention belongs to the field of medical biotechnology, and in particular relates to a method for detecting proteomes in colorectal cancer. Background technique [0002] The process of cancer metastasis involves a variety of cell biological behaviors, such as adhesion, movement, proliferation, and extracellular matrix degradation. Due to the complex pathological mechanism in the process of metastasis, quantitative proteomics technology is a powerful weapon to study the proteins involved in the metastasis and invasion of cancer cells. Proteomics technology can analyze the dynamic expression level of intracellular proteins with high invasiveness from an overall perspective, detect protein expression profiles in a panoramic manner, and mine early molecular events that occur during tumor metastasis, providing a new method for the study of complex mechanisms of cancer metastasis. train of thought. Transwell technology is an important strategy to esta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N27/62
CPCG01N33/57446G01N33/6848
Inventor 李斌胡会芳何庆瑜
Owner JINAN UNIVERSITY
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