A method to detect the proteome in colorectal cancer
A technology for proteomics and colorectal cancer, applied in the field of medical biology, can solve the problems of reducing the reliability of functional research results, and achieve the effects of avoiding heterogeneity, wide detection range and high detection accuracy
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[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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