Molecule mechanisms of TNF-α promoting the epithelial mesenchymal transformation of colon cancer cells through the CXCL10/CXCR3 signaling pathway
Objective To investigate the impact of TNF-α on epithelial mesenchymal transformation(EMT)and its molecular mechanism by regulating the expression of CXCL10/CXCR3 axis in colon cancer SW480 cells. Methods SW480 cells were cultured and treated with TNF-α and siRNA-CXCR3,respectively. The mRNA expression levels of CXCL10 and CXCR3 were detected using real-time PCR. Western Blot was performed to detect the expression levels of proteins involved in the CXCL10/CXCR3 pathway and EMT. Immunohistochemistry was used to observe changes in the expression of the EMT epithelial marker E-cadherin and the EMT interstitial marker vimentin. Transwell assay was conducted tp assess the migration and invasion abilities of cells in each group. Results The mRNA levels of CXCL10 and CXCR3 in the NF-α+si-CXCR3 group were higher than those in the NC group,and the difference was statistically significant(P<0.05). Additionally,the CXCR3 levels in the TNF-α+si-CXCR3 group were lower than those in the TNF-α group,and this difference was also statistically significant(P<0.05). The protein expression levels of CXCL10,CXCR3,vimentin and Fibronectin in the TNF-α group were higher than those in the NC group. However,the protein expression levels of E-CAD were lower than those in the NC group,and the difference was statistically significant(P<0.05). In the NF-α+si-CXCR3 group. The protein expression levels of CXCR3,vimentin and Fibronectin were lower than those in the TNF-α group. The protein level of E-CAD was higher than that in the TNF-α group. The expression of vimentin in the TNF-α group was higher than that of the NC group ,and the expression of E-cad was lower than that of the NC group. The differences were statistically significant(P<0.05). The expression of vimen-tin in the TNF-α+si-CXCR3 group was lower than that in the TNF-α group,and the difference was statistically significant(P<0.05). The expression of E-CAD was higher than that of the TNF-α group. The migration ability of the TNF-α group was higher than that of the NC group,and the difference was statistically significant(P<0.05). The cell migration ability of the TNF-α+si-CXCR3 group was lower than that of the TNF-α group,and the difference was statistically significant(P<0.05). The invasive ability of cells in the TNF-α group was higher than that of the NC group,and the difference was statistically significant(P<0.05). The cell invasion ability of the TNF-α+si-CXCR3 group was lower than that of the TNF-α group,and the difference was statistically significant(P<0.05). Conclusion TNF-α could induce epithelialization of colon cancer SW480 cells and promote their migration and invasion levels. This effect may be related to the activation of the CXCL10/CXCR3 signaling pathway.
万方数据
维普
