Effects Of DOT1L Silencing On The Proliferation,invasion And Migration Of Human Gastric Cancer Cells MGC-803 And Its Mechanism

Objective: To observe the effects of inhibiting DOT1 L gene on the proliferation,invasion and migration of gastric cancer cells MGC-803,and to explore its mechanism.Methods:(1)Western blot was used to detect the expression of DOT1 L in gastric cancer cell lines BGC-823,MGC-803 and SGC-7901,and cancer cell lines with high expression of DOT1 L gene were screened.(2)The viral vector plko-1puro-dot1l-sh RNA was constructed based on the DOT1 L target,and the gastric cancer cell lines with high expression of DOT1 L were selected for viral infection.(3)Real-time PCR was used to detect the expression level of DOT1 L m RNA in each group and to verify the interference efficiency.(4)CCK-8 assay was used to detect the effect of gene interference on the proliferation ability of gastric cancer cell lines,and the changes of colony formation ability after the expression of interfering gene were observed by colony cloning experiment(5)The migration and invasion ability were observed by Transwell experiment and cell scratch test respectively.(6)Western blotting was used to detect the expression of epithelial-mesenchymal-transition(EMT)related molecules(E-Cadherin,N-Cadherin,Vimentin)in cells after viral interference with the expression of DOT1 L gene.Results:(1)DOT1L was expressed in three gastric cancer cell lines at different degree,among which the DOT1 L expression was the highest in MGC-803 cells.Therefore,this study intends to select the gastric cancer mdc-803 cell line for subsequent experiments(2)sh RNA interfering DOT1 L gene expression model was successfully constructed.(3)The expression of DOT1 L m RNA and relevant protein was decreased significantly after inhibiting the expression of DOT1 L gene(P<0.05).(4)Compared with the control group,the ability of cell migration and invasion was significantly reduced(P<0.05).The expression of N-Cadherin and Vimentin protein was decreased in the cells(P<0.05)and the expression of E-Cadherin was increased(P<0.05).Conclusion: DOT1 L silencing affects the proliferation,migration and invasion abilities of gastric cancer cells MGC-803,and reverses the EMT process by regulating the expression of N-Cadherin,Vimentin,E-Cadherin,thereby reducing the migration and invasion ability of gastric cancer cells MGC-803.