miR-3612靶向SEMA4C调控肝细胞癌细胞的恶性生物学行为OA北大核心CSTPCD

Objective:To investigate the effect of miR-3612 targeting semaphorin(SEMA)4C on the proliferation and invasion ability of hepatocellular carcinoma cells.Methods:Forty pairs of cancerous tissues and corresponding paracancerous tissues of hepatocellular carcinoma that surgically resected at Yijishan Hospital,the First Affiliated Hospital of Wannan Medical College between May 2020 and September 2021 were collected for this study.Hepatocellular carcinoma Hep3B and Huh7 cells were routinely cultured and divided into control group,miR-3612 mimics-NC group,miR-3612 mimics group,miR-3612 inhibitor-NC group,miR-3612 inhibitor group,si-NC group,si-SEMA4C group,mimics-NC+pcDNA-NC group,miR-3612 mimics+pcDNA-NC group,and miR-3612 mimics+pcDNA-SEMA4C group.The corresponding nucleic acids and plasmids were transfected into each group of cells with transfection reagents.qPCR assay was used to detect the mRNA expression of miR-3612 and SEMA4C in hepatocellular carcinoma tissues and Hep3B and Huh7 cells.Dual-luciferase reporter gene assay and RNA immunoprecipitation assay(RIP)were used to validate the binding and regulatory relationship between miR-3612 and SEMA4C.qPCR and WB assays were used to detect the mRNA and protein expression of miR-3612 and SEMA4C in Hep3B and Huh7 cells after transfection in each group.The proliferation,migration and invasion abilities of Hep3B and Huh7 cells were detected by CCK-8 assay,cell scratch assay and Transwell assay,respectively.Results:miR-3612 was lowly expressed in hepatocellular carcinoma tissues and Hep3B and Huh7 cells(P<0.001),whereas SEMA4C was highly expressed(P<0.001).Overexpression of miR-3612 suppressed proliferation,migration,invasion,and expression of vimentin and SEMA4C proteins in Hep3B and Huh7 cells,promoted E-cadherin protein expression(P<0.05 or P<0.01 or P<0.001),and knockdown of miR-3612 promoted proliferation,migration,invasion and SEMA4C protein expression in Hep3B and Huh7 cells(P<0.05 or P<0.01 or P<0.001).Dual luciferase reporter gene assay and RIP assay confirmed that miR-3612 bound directly to SEMA4C(P<0.001),as indirectly evidenced by the negative correlation between miR-3612 and SEMA4C expression(P<0.001).Knockdown of SEMA4C significantly inhibited the proliferation,invasion and migration of Hep3B and Huh7 cells(P<0.05 or P<0.01 or P<0.001),and overexpression of SEMA4C reversed the inhibitory effect of miR-3612 overexpression on proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of Hep3B and Huh7 cells(P<0.05 or P<0.01 or P<0.001).Conclusion:miR-3612 affects the malignant biological behaviors of Hep3B and Huh7 cells by regulating SEMA4C expression,and it is expected to be a potential target for clinical hepatocellular carcinoma therapy.