Effects of overexpression of miR-193b-3p and PAK4 on proliferation,apoptosis,migration,and invasion of human colon cancer cells and their targeting relationships
Objective To investigate the effects of microRNA 193b-3p(miR-193b-3p)and P21-activated protein kinase 4(PAK4)overexpression on the proliferation,apoptosis,migration and invasion of human colon cancer cell line HCT116,and to verify the targeting relationship between them.Methods Human normal colon mucosal epithelial cells FHC and human colon cancer cells HCT116,SW480,LoVo were cultured in vitro,and the mRNA expression of miR-193b-3p and PAK4 was detected by RT-PCR.HCT116 cells in the logarithmic growth phase with good growth status were divided into four groups:miR-NC group(transfected with miRNA negative control),miR-193b-3p group(transfected with miR-193b-3p mimics),miR-193b-3p+PAK4-NC group(transfected with miR-193b-3p mimics + PAK4 empty vector),and miR-193b-3p+PAK4 group(transfected with miR-193b-3p mimics + PAK4 overexpression vector),respectively.The mRNA expression of miR-193b-3p and PAK4,the proliferation activity,apoptosis rate,the number of migrating cells and the number of invasive cells in each group were detected by RT-PCR,CCK-8 assay,flow cytometry,and Transwell experi-ment,respectively.The targeting relationship between miR-193b-3p and PAK4 was predicted by bioinformatics software Target Scan7.2,and verified by double luciferase reporter gene experiment.Results After overexpressing miR-193b-3p,the proliferation activity,migration and invasion abilities of HCT116 cells significantly decreased,the apoptosis rate significantly increased,and the expression level of PAK4 mRNA decreased(all P<0.05).Up-regulation of PAK4 expres-sion could partially reverse the inhibitory effect of overexpressing miR-193b-3p on proliferation,migration and invasion and the promotion of apoptosis of HCT116 cells(all P<0.05).The dual luciferase reporter gene assay showed that miR-193b-3p could target the 3'UTR region of PAK4 and inhibit its luciferase activity(both P<0.05).Conclusion Overex-pression of miR-193b-3p can inhibit the proliferation,migration and invasion of HCT116 cells,and promote their apopto-sis,and there is a targeting relationship between miR-193b-3p and PAK4.
microRNA-193b-3pcolon carcinomaHCT116 cellscell proliferationapoptosiscell migrationcell invasionP21-activated protein kinase 4
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