Effects of miR-203a-3p on invasion and migration of esophageal cancer cells and its targeted regulatory relationship with Survivin
Objective To explore the effects of miR-203a-3p on the invasion and migration of esophageal cancer cells and to analyze the targeted regulatory relationship with Survivin.Methods Esophageal cancer TE-1 cells and nor-mal esophageal mucosal epithelial HEEC cells in logarithmic growth phase were collected.The miR-203a-3p and Survivin mRNA were tested by RT-qPCR.The targeted regulatory relationship between miR-203a-3p and Survivin was analyzed by dual luciferase reporter assay.The TE-1 cells were divided into the control group,miR-203a-3p simulation group,miR-203a-3p inhibitory group,and negative control group,respectively.Cells in the miR-203a-3p simulation group were trans-fected with miR-203a-3p mimic,miR-203a-3p-inhibited group with miR-203a-3p inhibitor,and negative control group with miR-203a-3p NC,while cells in the control group were not transfected.Transfection efficiency was verified by RT-qP-CR.Cell viability was measured by MTT at 2,12,24,36 and 48 h after transfection,cell migration was measured by Scratch assay and cell invasion was measured by Transwell invasion assay.Results The mRNA expression of miR-203a-3p was lower and the mRNA expression of Survivin was higher in the esophageal cancer cells than in the normal esophageal mucosal epithelial cells(both P<0.05).TargetScan database analysis showed that the Survivin gene had seven consecu-tive nucleotide sequences at the 3′UTR end that could be complementary to miR-203a-3p.The result of dual luciferase re-porter assay showed that TE-1 cells co-transfected with Survivin-WT and plasmid-miR-203a-3p mimic plasmids had lower activity than other cells(P<0.05).The cell viability at 12,24,36 and 48 h was as follows:the miR-203a-3p inhibition group>the control group and the negative control group>the miR-203a-3p simulation group(all P<0.05).The cell mi-gration distance and the number of invasive cells were in the following order:the miR-203a-3p inhibition group>the con-trol group and negative control group>the miR-203a-3p simulation group(all P<0.05).Conclusion High expression of miR-203a-3p inhibits the migration and invasion of esophageal cancer cells,and the mechanism may be related to the tar-geted negative regulation of Survivin.
万方数据
维普
