An Experimental Study On The Regulation Of Proliferation And Invasion Of Gastric Cancer Cells By MIRNA-937-5p Targeting To RAB1A

Objective: To study the effect of Rab1 A targeted miR-937-5p on the proliferation and invasion ability of gastric cancer cells,which would provide theoretical basis for the early diagnosis of gastric cancer.Methods: Human gastric cancer cells SGC 7901,MGC-803 and normal gastric epithelial cells GES-1 were used in this study.Firstly,quantitative PCR was used to detect the expression levels of miR-937-5p and Rab1 A mRNA in these three cell lines.Furthermore,western blot was adapted to test the expression level of Rab1 A protein.Secondly,human gastric cancer MGC-803 cells were transfected with miR-937-5p mimics and miR-937-5p inhibitor.The expression level of Rab1 A mRNA was tested by quantitative PCR and protein expression level of Rab1 A was detected by western blot.In addition,the targeted region of miR-937-5p to the 3’-UTR region of Rab1 A gene was verified through dual-luciferase reporter assay.After transfection of human gastric cancer cells with Rab1 A plasmid and siRNA-Rab1 A,CCK-8 assay was used to detect changes in cell proliferation capacity.Flow cytometry was used to detect the changes in apoptosis level.Clone formation assay was used to measure changes in cell cloning ability and transwell was performed to detecte changes in cell invasion ability.mRNA expression levels of PCNA,MMP-2,MMP-9 and TIMP-1 were detected by quantitative PCR,and protein expression levels of PCNA,MMP-2,MMP-9 and TIMP-1 were detected by western blot.Results:1.According to the results of quantitative PCR and western blot detection,miR-937-5p was highly expressed in GES-1 cells,while its expression was relatively low in MGC-803 cells.Rab1 A was highly expressed in MGC-803 cells,but lowly in GES-1 cells.The expression level of Rab1 A in MGC-803 cells decreased with the increase of miR-937-5p(P<0.05),and vice versa.2.Dual-luciferase reporter gene assay was used to verify whether miR-937-5p could targeting the 3 ’-UTR region of Rab1 A gene.3.MGC-803 was successfully transfected with Rab1 A overexpressing plasmid.MTT,transwell and plate cloning showed that its enhanced proliferation,invasion,and clone formation(P<0.05),while flow cytometry showed no significant change in apoptosis level(P>0.05).4.MTT,transwell,plate cloning and flow cytometry displayed that RNAi to Rab1 A decreased proliferation,invasion,clone formation ability(P<0.05),and increased apoptosis(P<0.05).5.Overexpression of Rab1 A in gastric cancer cells could up-regulate the expression of PCNA,MMP2,MMP9 and down-regulate the expression of TIMP-1,while interference with Rab1 A had the opposite effect(P<0.05).Conclusions:1.Compared with GES-1 of normal gastric epithelial cells,miR-937-5p is low expressed in gastric cancer cells,while Rab1 A is low expressed in normal gastric epithelial cells GES-1.2.Rab1 A mRNA may be the target gene of miR-937-5p.3.Rab1 A can enhance the proliferation and invasion of MGC-803 cells.