Expression Of TGF-β₁ And TTG In Cisplatin-treated SW480 Colon Cancer Cells And Mechanism Research

Background:According to rough statistics,the incidence of colorectal cancer（CRC）ranks the third,its mortality ranks the fourth.In China,CRC is the third most common cancer,it is only lower than the rate of incidence of lung cancer and breast cancer.It is also the fifth most deadly cancer.Because the early symptoms of colon cancer are easily overlooked,many patients are diagnosed at an advanced stage,which greatly affects the treatment effect.Because cell proliferation,invasion and metastasis of colon cancer are very complex,it involves the interaction between multiple genes and pathways.Therefore,it is of great significance for improving the cure rate and quality of life on patients in CRC to reveal the changes of related factors on invasion,metastasis,reduce postoperative recurrence rate and assess prognosis.In the previous experiment,we have been working on colon cancer research and found that Transforming growth factor-β（TGF-β₁）and Tissue transglutaminase（tTG）may play an important role in colon cancer.However,its specific mechanism still need further investigation.TGF-β₁ is an important factor in cells,which regulates gene expression and affects cell processes on cell cycle distribution and tissue repair.In the early stage,TGF-β₁ is an important tumor suppressor and plays a role in inhibiting the proliferation of cancer cells.However,with the tumor gradually grows and infiltrates into the surrounding area,the overexpression of TGF-β₁ plays a role in promoting the invasion and metastasis of cancer cells to other tissues and organs.tTG is an important multifunctional enzyme in cells.As G protein,it can participate in various signal transduction in the cell.Some studies have confirmed that TGF-β₁ and tTG exist in the cytoplasm and nucleus to conduct cell signaling,and participate in importantphysiologicalprocesses,suchasinductionofcelladhesion,epithelial-mesenchymal transition（EMT）,cell growth and migration.Cisplatin（DDP）is the first-line drug for clinical combination chemotherapy.Cisplatin can promote the cross-linking between chlorine and DNA after dissociation,and destroy DNA replication efficiency.Then,the expression of TGF-β₁ and tTG in cisplatin-treated SW480 cells and the mechanism research had become the focus in our experiments.Purpose:In this experiment,the expression of TGF-β₁ and tTG in colon cancer and paracancerous tissue was detected in order to elucidate the role of TGF-β₁ and tTG in colon cancer tissue.The survival rate and the distribution of the cycle of SW480 cells,as well as the changes of TGF-β₁ and tTG expression was detected after 24-hour incubation of cisplatin to elucidate the targeting effect in cisplatin-treated SW480cells.The TGF-β₁ inhibitor and inducer were used to treat SW480 cells for 24 hours.Then the expression of tTG in SW480 cells was analyzed by Western blot and immunofluorescence to explore the possible regulatory mechanisms between this two protein.Method:1.HE staining confirmed that the specimens obtained were colon cancer and paracancerous tissues,which lay a foundation for the diagnosis of patients’condition.2.The expression of TGF-β₁ and tTG was analyzed by Immunohistochemical staining and Western Blot in colon cancer tissue.3.The survival rate of SW480 cells treated with cisplatin was determined by MTT assay.4.The changes of the periodic distribution of SW480 cells treated with cisplatin were analyzed by Flow cytometry.5.The expression levels of tTG and TGF-β₁ in cisplatin-treated SW480 cells were analyzed by immunofluorescence and Western Blot.6.The expression of tTG and TGF-β₁ in SW480 cells treated with TGF-β₁inhibitor and inducer was analyzed by immunofluorescence and Western Blot.Results:1.HE staining showed that the selected pathological tissue was colon cancer tissue;Immunohistochemistry results showed that the expression of tTG and TGF-β₁in colon cancer tissue increased significantly.2.Compared with the control group,the survival rate of SW480 cells decreased after cisplatin treatment.3.Compared with the control group,cisplatin could regulate the cycle in SW480cells.4.Compared with the control group,the expression of tTG and TGF-β₁ in cisplatin-treated SW480 cells was decreased.5.Compared with the control group,the expression of tTG and TGF-β₁ might increase after 20 ng/mL TGF-β₁ inducer treatment in SW480 cells.6.Compared with the control group,TGF-β₁ inhibitor could inhibite the expression of tTG and TGF-β₁ in SW480 cells.Conclusion:1.TGF-β₁ and tTG played an important role in colon cancer.2.Cisplatin could inhibit the survival rate of SW480 cells and regulate the cycle of SW480 cells.3.Cisplatin could inhibit the expression of tTG and TGF-β₁ in SW480 cells.4.Abnormalities in TGF-β₁ might regulate the expression of tTG.