KIF23基因沉默对肝癌HepG2细胞增殖、迁移及侵袭的影响  

Effects of KIF23 Gene Silencing on Proliferation,Migration and Invasion of Human Hepatocellular Carcinoma HepG2 cells

作　　者：刘素娟 林曲[2] 柏明军[1] 周楚人 陈俊伟[1] 吴春[1] 黄明声[1] 

机构地区：[1]中山大学附属第三医院介入血管外科,广东广州510630 [2]中山大学附属第三医院肿瘤内科,广东广州510630

出　　处：《中山大学学报（医学版）》2018年第1期34-40,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金面上项目(S2012010008705);广东省自然科学基金博士启动项目(2015A030310171)

摘　　要：【目的】研究KIF23基因对肝癌HepG2细胞体外增殖、迁移及侵袭的影响,并探讨可能的作用机制。【方法】化学合成针对KIF23基因的siRNA,通过脂质体转染至HepG2细胞中,应用qRT-PCR和Western blot法检测转染后HepG2细胞中KIF23 mRNA和蛋白表达变化。采用CCK-8法和平板克隆法检测KIF23沉默后对HepG2细胞增殖的影响,并通过细胞划痕实验和Transwell侵袭实验分别检测KIF23沉默后对迁移和侵袭能力的影响。Western blot法检测KIF23基因沉默后对蛋白激酶B(PKB/Akt)及磷酸化Ak(tp-Akt)蛋白表达的影响。【结果】KIF23-siRNA能有效沉默HepG2细胞中KIF23的mRNA和蛋白表达(P值均<0.01)。CCK-8实验、平板克隆实验、划痕实验和Transwell侵袭实验结果表明,KIF23-siRNA2干扰组与阴性对照组、空白对照组比较,细胞的增殖、迁移和侵袭能力均明显受到抑制(P值均<0.05)。KIF23-siRNA2干扰组HepG2细胞中总Akt蛋白的表达水平未发生变化,但磷酸化Akt蛋白的表达水平明显下调(P<0.05)。【结论】KIF23可能通过激活Akt信号转导通路促进人肝癌细胞的增殖、迁移及侵袭能力,KIF23有望成为肝癌基因治疗的新靶点。【Objective】To investigate the effect of KIF23 gene expression on the proliferation,migration and invasion of human hepatocellular carcinoma HepG2 cells in vitro,and to explore the possible mechanism.【Methods】The KIF23 si RNA was transfected into HepG2 cells by lipofectamine 3000. The expression of KIF23 mRNA and protein in HepG2 cells was detected by qRT-PCR and Western blot. The effect of silencing KIF23 on the proliferation of HepG2 cells was studied by CCK-8 assay and plate clone formation assay. The tumor cell abilities of migration and invasion after transfection were measured by scratch assay and Transwell assay. The expression of protein kinase B(PKB/Akt)and phosphorylated Akt(p-Akt)protein in HepG2 cells transfected with KIF23-siRNA2 was detected by Western blot.【Results】KIF23-siRNA could effectively silence the expression of KIF23 mRNA and protein in HepG2 cells(P<0.01).The results of CCK-8 assay,plate clone formation assay,scratch assay and Transwell assay demonstrated that the cell proliferation,migration and invasion ability of the KIF23-siRNA2 interference group were significantly inhibited,compared to the negative control group and the blank control group(P<0.05). The expression level of total Akt protein in HepG2 cells was not changed,but the expression level of phosphorylated Akt protein was down-regulated(P<0.05).【Conclusions】KIF23 may promote the proliferation,migration and invasion of human hepatocellular carcinoma cells by activating Akt signal transduction pathway. KIF23 is expected to be a new target for gene therapy of hepatocellular carcinoma.

关 键 词：肝细胞癌 RNA干扰 KIF23 细胞增殖 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]