Effects of RNAi-mediated SSBP1 gene silencing on proliferation,invasion and metastasis of human hepatocellular carcinoma HepG2 cells in vitro

MA Rongfen; CHEN Qiuying; LIU Yanfen

doi:10.3969/j.issn.1002-7386.2017.17.004

Abstract：Objective To investigate the effects of RNAi-mediated SSBP1 gene silencing on proliferation,invasion and metastasis of human hepatocellular carcinoma HepG2 cells in vitro.Methods The high effective targeting SSBP1 siRNA was obtained by construction and screening,which was transfected into HepG2 cells by liposomes mediated transient transfection.The cells were divided into 3 groups: negative control group,blank transfection group (blank control group) and transfection group.The expression levels of SSBP1 mRNA and protein were detected by Real time-PCR and Western-Blot after the transfection.CCK-8 method was used to detect cell proliferation.Flow cytometry was used to detect the cell cycle,cell apoptosis and mitochondrial trans-membrane potential.The scuffing test and Transwell invasion assay were used to detect invasion and metastasis ability of cells.Moreover the expressions of gene and protein related with cell proliferation,invasion and metastasis were detected by Western Blot.Results The SSBP1 siRNA could obviously inhibit the expressions of SSBP1 mRNA and protein.As compared with negative control group and blank control group,after HepG2 cells were transfected by SSBP1 siRNA,the cell proliferation ability, cell proportion at phase G2 and phase S, mitochondrial trans-membrane potential were obviously decreased,however, the cell proportion at phase G1 and cell apoptosis rate were significantly increased (P<0.05).Meanwhile the expression levels of PCNA,Bcl-2 and MMP-9 proteins were obviously down-regulated,however, the expression levels of Bax proteins were significantly up-regulated (P<0.05).Conclusion The targeting silencing of SSBP1 gene can inhibit proliferation,invasion, metastasis of HepG2 cells,and can induce cell apoptosis via mitochondria pathway.