Experimental Study On Apoptosis Of Peritoneal Mesothelial Cells Induced By Colorectal Cancer Microenvironment

Objective:Colorectal cancer is one of the most common malignant tumors in the world.It is the third and fourth most common malignant tumors in the world that cause cancer-related death.With the continuous improvement of people’s living standards and changes in eating habits,China’s knots the incidence and mortality of rectal cancer are on the rise.At the same time,about 25% of patients with colorectal cancer have distant metastasis at the time of diagnosis.The peritoneal metastasis is second only to liver metastasis,ranking second in the metastatic site.Because China has not implemented and promoted colorectal cancer screening.Measures,therefore,the incidence of peritoneal metastasis of colorectal cancer in China is higher than that of other developed countries.Therefore,the importance of peritoneal metastasis is important to find out the mechanism of peritoneal metastasis of colorectal cancer and related inflammatory mediators.It is important in the peritoneal metastasis of colorectal cancer.There may be some application value in the treatment,which not only can prolong the survival of the patient,but also help to improve the prognosis of the patient.Method: 1.The supernatant secreted by colorectal cancer cells was collected and co-cultured with human peritoneal mesothelial cell line HMr SV5 as an experimental group;serum-free medium and peritoneal mesothelial cells HMr SV5 were co-cultured as a control group.(1)Using an inverted microscope to observe changes in the morphology of mesothelial cells(2)Detection of proliferation of mesothelial cells using CCK-8 assay(3)Flow cytometry and fluorescent staining to determine the proportion of mesothelial apoptosis(4)Immunoblotting was used to detect the expression of apoptosis-related proteins Bcl-2 and Bax 2.peritoneal mesothelial cells HMr SV5 and colorectal cancer cells co-cultured through Transwell chambe Observe whether damaged peritoneal mesothelial cells have chemotactic effects on colorectal cancer cells.Result: 1.Colorectal cancer cell supernatant changes the morphology of peritoneal mesothelial cells After exposure to colorectal cancer,the peritoneal mesothelial cells are largely detached and become round and cylindrical.2.Colorectal cancer supernatant inhibits proliferation of peritoneal mesothelial cells The serum-free medium and the colorectal cancer supernatant were separately added into the peritoneal mesothelial cells,and the proliferation ability of the cells was detected by CCK-8.Compared with the serum-free medium,the proliferation of the peritoneal mesothelial cells added to the colorectal cancer supernatant was obvious.reduce.3.Colorectal cancer cell supernatant promotes apoptosis of peritoneal mesothelial cells Apoptosis was detected by Annexin V-FITC.The results showed that the supernatant of colorectal cancer cells promoted the apoptosis of peritoneal mesothelial cells.The anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax were detected by immunoblotting.The expression of the colorectal cancer cell supernatant significantly inhibited the expression of the anti-apoptotic protein Bcl-2 in mesothelial cells and increased the expression of the pro-apoptotic protein Bax.4.damaged residual mesothelial can be reversed for colorectal cancer cells,making their migration and transfer power increased Colorectal cancer cells and mesothelial cells were co-cultured in Transwll chamber,and colorectal cancer cells were cultured alone as a control.As a result,the migration ability of colorectal cancer cells was significantly improved after co-culture with mesothelial cells.Conclusion: 1.Colorectal cancer cells can inhibit mesothelial cell proliferation through their secretions and induce apoptosis in mesothelial cells.2.Mesothelial cells stimulated by injury can counteract colorectal cancer cells and promote their migration.