| Objectives: To preliminarily investigate the mechanism and effect of miR-924 on the proliferation of gastric cancer MGC803 cells and provide experimental basis for further understanding the molecular mechanism of gastric cancer.Methods: 1.The microRNAs targeting TGM2 gene were predicted and analyzed by bioinformatics software.The expression of miR-924 was detected by qRT-PCR in gastric cancer MGC803 cells.2.The 3’-UTR binding site of TGM2 Gene regulated by miR-924 was predicted by Targetscan software.Targeted binding of miR-924 to TGM2 was detected by Luciferase detection.3.The expression of TGM2 was detected in MGC803 cells introduced into miR-924.The effect of miR-924 on the proliferation of MGC803 was observed by growth curve,plate cloning and soft agar colony formation experiments.4.The expression of JAK3,STAT3 and their phosphorylated proteins was detected by Western-blotting in MGC803 cells.Results:1.The results of bioinformatics software predicted showed that the TGM2 gene targeting by miR-924.The results of qRT-PCR showed that the expression of miR-924 was low in gastric cancer MGC803 cells(P<0.05).2.miR-924 binds to 864-871 nucleotides of TGM2 gene 3’UTR.The luciferin reporter activity of 3’UTR sequence containing TGM2 gene was decreased after the overexpression of miR-924,and the expression of TGM2 gene and protein was decreased in MGC803 cells(P<0.05).3.The growth rate of MGC803 cells was slowed down,the volume and number of clones of plate cloning and soft agar colony formed was decreased,and the proliferation ability of cells was decreased after the overexpression of miR-924.4.The results of Western-blotting showed that the expression of JAK3 and STAT3 proteins was no significant difference in in MGC803 cells after overexpression of miR-924,but the phosphorylation level of both decreased(P<0.05).Conclusions: miR-924 targetedly inhibit the expression of TGM2,down-regulate the activation of JAK3/STAT3 pathway and reduce the proliferation of MGC803 cells. |
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