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

作　　者：蒋清虎 马吉安[1] 谭孝华[1] 吴忠均[2] 

机构地区：[1]达州市中西医结合医院肝胆外科,达州635000 [2]重庆医科大学附属第一医院肝胆外科,重庆400016

出　　处：《郑州大学学报（医学版）》2016年第6期723-726,共4页Journal of Zhengzhou University(Medical Sciences)

摘　　要：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.

关 键 词：肝癌 侵袭 ARK5基因 RNA干扰 SMMC-7721细胞 

分 类 号：R735.7[医药卫生—肿瘤]