Effect Of SOD3 Inhibited By MicroRNA-339-3p On Proliferation Of Immortalized Gastric Epithelial Cells GES-1

Objectives: To preliminarily investigate the effect of microRNA-339-3p on the proliferation of immortalized gastric epithelial cells GES-1 and its mechanism,so as to provide experimental basis for further understanding the molecular mechanism of gastric cancer.Methods: 1.Firstly,the microRNAs of SOD3 gene interaction were analyzed by online bioinformatics software,and then the expression of microRNA-339-3p in GES-1 cells was detected by real-time fluorescence quantitative PCR.2.The interaction sites between microRNA-339-3p and the 3 ’-UTR domain of SOD3 gene were analyzed by bioinformatics software,and the interaction between microRNA-339-3p and the 3’-UTR domain of SOD3 gene was analyzed by luciferase reporter gene.3.microRNA-339-3p was transfected into GES-1 cells,and the expression of SOD3 was detected by real-time fluorescence quantitative PCR and Western blotting in GES-1 cells.4.The effects of microRNA-339-3p on the proliferation of GES-1 cells were observed by cell growth curve and plate cloning.5.The activation of JAK3/STAT3 signaling pathway was detect by Western-blotting.Results:1.Through the analysis of online bioinformatics software,it was found that microRNA-339-3p interacted with SOD3 gene.The results of real-time fluorescent quantitative PCR showed that microRNA-339-3p was low expression in GES-1 cells(P < 0.05).2.Bioinformatics software analysis showed that microRNA-339-3p interacted with the385-392 nucleotide of 3’UTR of SOD3 gene.The luciferase reporter gene analysis showed that the 3’UTR mutation of SOD3 gene was cotransfected with microRNA-339-3p mimics,anti-microRNA-339-3p or mir-control into GES-1 cells,but the luciferase activity of the three genes was not different,but the 3’UTR sequence of SOD3 gene and microRNA-339-3p were co transfected The luciferase activity of GES-1 cells co transfected with microRNA-339-3p mimics,anti-microRNA-339-3p or microRNA-control was significantly reduced(P< 0.05).3.The results of real-time fluorescent quantitative PCR and Western blotting showed that after microRNA-339-3p transfection,the mRNA and protein expression of SOD3 in GES-1 cells decreased(P< 0.05).4.The results of cell growth curve showed that after the high expression of microRNA-339-3p,the growth speed of GES-1 cells was significantly accelerated(P< 0.05).The results of plate clone formation experiment also showed that after the high expression of microRNA-339-3p,GES-1 cells had a large volume and a large number of clones(P< 0.05).5.The results of Western blotting showed that the activation of JAK3/STAT3 signaling pathway in GES-1cells was promoted by overexpression of microRNA-339-3p(P< 0.05).Conclusions: microRNA-339-3p down-regulates the expression of SOD3,promots the activation of JAK3/STAT3 signaling pathway and enhanced the proliferation of GES-1 cells.