ABSTRACT
Objective:To investigate the effects of miR-195 on the migration, invasion and epithelial mesenchymal transition (EMT) of human gastric cancer cell line (HGC-27) and its mechanism.Methods:HGC-27 cells were cultured in vitro and they were randomly divided into control group, miR-195 negative control (NC) group and miR-195 mimics (mimics) group; the expressions of miR-195 and SALL4 mRNA were detected by real-time fluorescence quantitative PCR (RT-qPCR) ; The changes in cell morphology of each group were observed under a microscope; the expressions of SALL4, E-cadherin, N-cadherin and Vimentin in HGC-27 cells were detected by Western blot; MTT method was used to detect the change of HGC-27 cell survival rate; the abilities of migration and invasion in HGC-27 cells were detected by scratch test and invasion test (Transwell) ; and the targeting relationship between miR-195 and SALL4 was confirmed by double Luciferase Report.Results:Compared with those in control group and miR-195 NC group, the miR-195 expression level, apoptosis rate and E-cadherin protein expression level of HGC-27 cells in miR-195 mice group were significantly higher ( P<0.05) , the cell survival rate, scratch healing rate, invasion number, SALL4 mRNA and SALL4, N-cadherin, Vimentin protein expression levels were significantly lower ( P<0.05) , the morphology of HGC-27 cells returned to normal, and the epithelial mesenchymal behavior was significantly reduced; in addition, there was a binding site between miR-195 and SALL4 mRNA 3'UTR region, and compared with that in SALL4-3'UTR-WT + miR-195 NC group, the luciferase activity of SALL4-3'UTR-WT + miR-195 MICs group was lower ( P<0.05) . Conclusion:MiR-195 may inhibit the migration, invasion and epithelieal-mesenchymal transition of human gastric cancer cells by targetingly inhibiting the expression of SALL4.
ABSTRACT
Sal-like 4 (SALL4) plays an important role in promoting the cellular proliferation and maintaining the pluripotency of embryonic stem cells and tumor cells.In fully differentiated cells,the expression of SALL4 is silenced or down-regulated.However,the expression of SALL4 is found to be restored or up-regulated in a variety of non-germ cell tumors.Besides,the expression of SALL4 is often associated with disease progression,treatment effect and prognosis.Therefore,examining the expression level of SALL4 will be of great importance in the diagnosis of disease and monitoring the disease progression for non-germ cell tumors.