siRNA沉默ARK5基因对SMMC-7721细胞侵袭及MMP-2蛋白分泌的影响
Effect of siRNA silencing ARK5 gene on invasion and secretion of MMP-2 of SMMC-7721 cells

目的:观察siRNA沉默ARK5基因的表达对肝癌SMMC-7721细胞侵袭及MMP-2蛋白分泌的影响。方法:采用qRT-PCR和Western blot法检测SMMC-7721细胞和正常肝细胞LO2中ARK5的表达。体外合成靶向ARK5基因的siRNA,转染肝癌SMMC-7721细胞,并设阴性对照和空白对照,采用Western blot方法检测3组细胞中ARK5蛋白的表达,通过Transwell 小室模型检测细胞的体外侵袭能力,采用ELISA法检测细胞培养基中MMP-2蛋白的含量。结果:SMMC-7721细胞中ARK5 mRNA和蛋白的表达水平高于LO2细胞(P＜0.05); 沉默ARK5基因后,SMMC-7721细胞的侵袭受到抑制,且MMP-2蛋白的分泌水平低于阴性对照和空白对照(P＜0.05)。结论:沉默ARK5基因可能通过MMP-2途径抑制SMMC-7721细胞的侵袭。
Aim: To study the effect of ARK5 gene silencing on the invasion of SMMC-7721 cells.Methods: The expression of ARK5 was determined by qRT-PCR and Western blot in SMMC-7721 and LO2 cells,respectively. ARK5 siRNA was constructed and transfected into SMMC-7721 cells, and negative group and blank group were set up; the expression of ARK5 protein was determined by Western blot,and the invasion of cells in 3 groups were examined by Transwell assay. MMP-2 protein in culture medium was determined by ELISA.Results: The mRNA and protein expression levels of ARK5 in SMMC-7721 cells were significantly higher than those in LO2 cells(P＜0.05). After ARK5 siRNA transfection, the invasion of SMMC-7721 cells was significantly inhibited,and MMP-2 protein secreted by the cells was significantly lower than those in negative group and blank group(P＜0.05).Conclusion: ARK5 silencing could effectively inhibit invasion of SMMC-7721 cells via MMP-2 pathway