Inhibitory Effect Of MiR-200c On TGF-?1-induced Epithelial-mesenchymal Transition Of Gastric Carcinoma Cell Line

ObjectiveTo determine whether TGF-?1 can induce EMT in gastric carcinoma;In the course of EMT,the relationship between TGF-?1 and miR-200 c was studied.Finally,the correlation between the expression of miR-200 c and EMT induced by TGF-?1 was observed.Mehtods1.To induce gastric carcinoma cells at different time with the same concentration of TGF-?1(10ng/ml).Western blot was used to detect the expression of EMT related marker protein,CCK-8 was used to detect cell proliferation;2.After establishing EMT model,quantitative PCR was used to detect the effect of TGF-?1 on miR-200 c expression;3.miR-200 c was overexpressed by liposome transient transfection and gastric cancer cell lines were induced by TGF-?1.The morphology of gastric cancer cells was observed by microscope,and the expression of EMT related markers was detected by Western blot.Results1.To induce gastric cancer cells for 0h,12 h,24h,36 h with the same concentration of TGF-?1(10ng/ml),then Western blot showed that the expression of E-cadherin decreased with time,while vimentin increased with time,the results of CCK-8 showed that the proliferation ability of TGF-?1 treated group was also higher than that of the control group at 0 h,12 h,24 h and 36 h,so EMT cell model of gastric cancer in vitro is established.2.After establishing EMT model,the results of quantitative PCR indicated that the expression of miR-200 c in TGF-?1 group was lower than that in negative control group.3.To increase the expression of miR-200 c in gastric carcinoma cells and induce them with TGF-?1,observation of cell morphology under microscope shows that:Compared with the TGF-?1 treated group,the cell morphology was reversed and tended to the connom SGC-7901 cell strain in TGF-?1+miR-200 c group;western blot showed that:to compare TGF-?1 treatment group and TGF-?1+miR-200 c group,the expression of E-cadherin in TGF-?1+miR-200 c group was relatively increased,and vimentin was relatively decreased.ConclusionsTGF-?1 can induce EMT in gastric carcinoma,epithelial cells transformed into stromal cells with stronger proliferative ability;In EMT of gastric carcinoma,TGF-?1 can down regulate the expression of miR-200c;the over expression of miR-200 c can inhibit the EMT process induced by TGF-?1...