MiR-10a-5p Promotes The Invasion And Migration Of Gastric Cancer Cells By Targeting RORA

Objective:To explore whether miR-10a-5p affects the invasion and migration of gastric cancer cells and the mechanism of invasion and migration by targeting the retinoic acid receptor-related orphan receptor A(RAR-related orphan receptor A,RORA).Methods:qRT-PCR was used to detect the expression levels of miR-10a-5p in human normal gastric mucosa cells GES-1 and different gastric cancer cell lines AGS、SGC-7901、BGC-823.The gastric cancer cell line SGC-7901 with high expression of miR-10a-5p was used as a follow-up experiment to study the gastric cancer cell line.The experiment was divided into Control group(trasfection of Lipofectamine~?2000),NC group(Lipofectamine~?2000 and NC Sequence,transfection irrelevant sequence)and miR-10a-5p inhibitor group(Lipofectamine~?2000 and miR-10a-5p inhibitor),qRT-PCR was used to detect the expression level of miR-10a-5p in each group of cells after transfection.The bioinformatics website Target Scan predicted the targeted binding site of miR-10a-5p and RORA.qRT-PCR and Western Blot experiments analyzed the relationship between miR-10a-5p and RORA.Colony formation and Transwell experiments were used to detect the effects of down-regulation of miR-10a-5p on the proliferation and invasion and migration ability of SGC7901 cells.The experiment was divided into Control group(transfection of Lipofectamine~?2000 only),RORA NC transfection group(transfection of Lipofectamine?2000 and NC sequence),RORA OE group(Lipofectamine?2000 and RORA OE),transwell experiment detected the effect of up-regulation of RORA on the invasion and migration ability of SGC-7901 cells.Western Blot method detected the relative expression levels of invasion and migration-related proteins E-cadherin,N-cadherin and Snail.Results:miR-10a-5p was highly expressed in gastric cancer cell lines when compared to normal gastric mucosal cells(P<0.05).Bioinformatics software Target Scan predicted that RORA 3’UTR has potential sites for miR-10a-5p targeted binding(P<0.05),and miR-10a-5p can negatively regulate RORA gene expression.After down-regulation of miR-10a-5p,the proliferation,invasion and migration of SGC-7901 cells were significantly inhibited(P<0.05).Overexpression of RORA could inhibit the invasion and migration ability of SGC-7901 cells(P<0.05).N-cadherin and Snail protein expression decreased,and E-cadherin expression increased(P<0.05).Conclusion:miR-10a-5p can promote the invasion and migration of gastric cancer cells by targeting RORA,and the mechanism of invasion and migration may be related to epithelial-mesenchymal transition.