Mechanism Of Mitochondrial TRAP1 Molecule In Intestinal Cancer Metastasis

Objective: To study the tumor necrosis factor receptor-related protein 1(TRAP1)to promote the epithelial-mesenchymal transition process of intestinal cancer.By examining the effects of TRAP1 on the biological characteristics of human and in vitro and intestine cancer cells,the mechanism of TRAP1 in the development of intestinal cancer was elucidated.Methods : Transfection was used to knock down and increase the expression of TRAP1 in intestinal cancer cells.MTT,Transwell,scratch,plate cloning,flow cytometry and other experimental techniques were used to detect the related functions and biological characteristics of intestinal cancer cells.Immunofluorescence assay was used to determine the localization of TRAP1 in intestinal cancer cells and the localization and expression of TRAP1 in intestinal cancer cells after lentiviral packaging.Real-time quantitative PCR,Western blot and immunofluorescence experiments were used to detect down-regulation.The expression levels of TRAP1 gene in the intestinal cancer cell lines HCT116 and SW620 after TRAP1 and the intestinal cancer cell lines HCT116 and HCF-8 over expressing TRAP1,and the expression levels of EMT markers at mRNA and protein levelsand their changes.Results: Intestinal cancer cells treated with lentiviral packaging were selected and revealed by fluorescent labeling.The expression of RAP1 was mainly located around the nucleus;the down-regulated TRAP1 expression of intestinal cancer cell lines HCT116 and SW620 was decreased,and TRAP1 over expressed intestinal cancer.The proliferation and colony forming ability of HCT116 and HCF-8 were enhanced.The knockdown of TRAP1 decreased the migration and invasion ability of intestinal cancer cell lines.Over expression of TRAP1 enhanced the migration and invasion of intestinal cancer cell lines.Down-regulation of TRAP1 intestinal cancer The apoptotic rate of the cell line was increased,and the apoptosis rate of the intestinal cancer cell line with up-regulated TRAP1 was decreased.The expression of EMT-related gene in the lentivirus-packaged intestinal cancer cell line was correlated with the expression of TRAP1,that is,TRAP1 interfered with the intestinal cancer cell line HCT116.EMT-related protein expression levels were normal in the control group,E-Cadherin expression was up-regulated and Vimentin expression was down-regulated.At the same time,the expression of E-Cadherin was also up-regulated in the expression of TGF1-interfering intestinal cancer cell line SW620 compared with the normal control group,while the expression level of EMT-related protein in TRAP1over-expressing intestinal cancer cell HCF-8 was compared.In the normal control group,the expression of E-Cadherin was down-regulated.Conclusions: TRAP1 is relatively high expression in intestinal cancer cell lines HCT116 and SW620,and relatively low expression in intestinal cancer cell line HCF-8;TRAP1 inhibits apoptosis of intestinal cancer cell line and promotesproliferation of intestinal cancer cells The ability of invasion,metastasis and clonal formation;TRAP1 affects the EMT process of intestinal cancer cells,induces EMT in intestinal cancer cells,promotes invasion and distant metastasis of intestinal cancer cell lines,and TRAP1 can be used as a potential biomarker for intestinal cancer.Screen for candidate genes.