Expression Of Twist2 In Gastric Cancer Cells And Its Effect On Epithelial-mesenchymal Transition

Objective: Twist2 mRNA and protein expression in human gastric cancer cell lines were detected.Gastric cancer cell lines were transfected with shRNA-encoded plasmids to suppress the expression of Twist2.The expression of epithelial-mesenchymal transition-related factors cadherin E(E-cadherin)and vimentin(Vimentin)in gastric cancer cells before and after transfection was analyzed.To investigate the role of Twist2 in epithelial-mesenchymal transition(EMT)of gastric cancer cells,and to observe the effect of Twist2 on the invasion and migration ability of gastric cancer cells.Methods: 1.Culture gastric cancer SGC-7901,BGC-823,and MGC-803 cell lines.After extracting mRNA and protein,q-PCR and Western blot were used to detect the expression levels.2.Select the BGC-823 cell line with the highest expression level of Twist2 mRNA and protein for further research.The gastric cancer BGC-823 cell line was transfected with a shRNA-encoded plasmid.The experiment was divided into three groups: Blank group;Twist2 negative control group(NC-sh);Twist2 silent group(Twist2-sh).Q-PCR and Western blot were used to detect the expression of Twist2 mRNA and protein in the three groups of cells,and the expression of EMT-related factors E-cadherin and Vimentin in each group of cells.3.Transwell chamber invasion and migration experiments were used to detect the invasion and migration ability of cells in each group.Results: 1.Expression of Twist2 mRNA and protein in human gastric cancer cell lines SGC-7901,BGC-823,and MGC-803,and the expression level in BGC-823 cell line was significantly higher than other cell lines(P <0.01).2.(1)q-PCR was used to detect the expression of Twist2 mRNA in each group of cells.The results showed that compared with the Twist2 silent negative control group(NC-sh),the expression of Twist2 mRNA in the Twist2 silent group(Twist2-sh)was significantly reduced(P < 0.01).(2)Western blot detected the expression of Twist2 protein in each group of cells.The results showed that compared with the NC-sh group,the expression levels of E-cadherin,Vimentin and Twist2 in the Twist2-sh group were changed,and the differences were statistically significant(P < 0.01).The expression of Twist2 protein was significantly down-regulated,the epithelial marker E-cadherin increased with the decrease in Twist2 expression,and the interstitial marker Vimentin decreased with the decrease in Twist2 expression.3.The invasion and migration ability of human gastric cancer cell line BGC-823 Twist2 silencing group was significantly weakened.Conclusions: 1.Expression of Twist2 mRNA and protein in a variety of human gastric cancer cell lines(SGC-7901,MGC-803,BGC-823);2.shRNA-encoded plasmid can significantly inhibit Twist2 mRNA and Protein expression level.Down-regulation of Twist2 can increase the expression of E-cadherin and decrease the expression of Vimentin,which proves that Twist2 can promote the EMT process of human gastric cancer cells.3.Twist2 can increase the invasion and migration capacity of human gastric cancer cells through EMT.