沉默Rock2对肝癌细胞增殖及凋亡作用的研究  

Study of Rock2 on the Proliferation and Apoptosis of Hepatocellular Carcinoma Cells

作　　者：刘天德[1] 袁荣发[1] 王庆诺[1] 蒋成行[1] 杨志强[1] 邵江华[1,2] 

机构地区：[1]南昌大学第二附属医院肝胆外科,330006 [2]江西省分子医学重点实验室

出　　处：《天津医药》2012年第10期979-982,共4页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:30860272);江西省研究生创新基金资助项目(项目编号:YC10A018)

摘　　要：目的:探讨沉默Rock2基因对人肝癌细胞Huh-7和HepG2增殖和凋亡作用的影响。方法:实验分为空白对照组、干扰无意义组、转染PBS组及干扰Rock2组。将Rock2干扰质粒shRock2转染到人肝癌细胞Huh-7和HepG2中,通过实时荧光定量PCR检测Rock2mRNA的表达水平;Westernblot检测Rock2蛋白的表达水平;MTT比色法检测沉默Rock2后对Huh-7和HepG2细胞增殖抑制的影响;流式细胞术检测沉默Rock2对Huh-7和HepG2细胞周期及早期凋亡的变化。结果:将shRock2转染肝癌细胞系Huh-7和HepG2后,Rock2mRNA以及Rock2蛋白的表达水平均明显下降。沉默Rock2表达后,MTT结果显示Huh-7和HepG2细胞较对照组细胞增殖能力明显减弱(P<0.01);Huh-7和HepG2细胞G0/G1期细胞比例明显升高,而S期和G2/M期细胞比例明显降低,与对照组比较差异有统计学意义(P<0.01);肝癌细胞的早期凋亡较对照组明显增加(P<0.01)。结论:沉默Rock2能明显抑制肝癌细胞系Huh-7和HepG2的增殖,并诱导其早期凋亡,提示Rock2可作为肝癌基因治疗的一个新的分子靶点。Objective： To investigate the effects of Rock2 on proliferation and apoptosis of hepatocellular carcinoma Huh-7 and HepG2 cell hnes. Methods： There were four experimental groups in this study,including untreated group, non- targeting group, PBS group and shRock2 group. Hepatocellular carcinoma Huh-7 and HepG2 cell lines were transfected with shRock2. Reverse transcriptase polymerase chain reaction （RT-PCR） and Western blot assays were used to detect the expression levels of Rock2 mRNA and protein, respectively. The cell proliferation of Huh-7 and HepG2 was measured by MTT assay. The ceil cycle was detected by PI staining method through flow cytometry and the earlier apoptosis was demonstrated by Annexin V apoptosis kit. Results： Results of RT-PCR and Western blot showed that the expression levels of Rock2 mRNA and protein were significantly decreased in shRock2 transfected Huh-7 and HepG2 cells. After the silence of Rock2, the cell proliferation was blocked in the shRock2 ceils compared to that of control group （P 〈 0.01）. The cell- cycle arrest was promoted at the G0/G1 phase. But the percentage of ceils in S phase and G2/M decreased compared to that of control groups （P 〈 0.01）. The earlier apoptotie rate of shRock2 Huh-7 and HepG2 cells was significantly increased compared with that of control group （P 〈 0.01）. Conclusion： After Rock2 silencing, the proliferation was significantly inhibited and early apoptosis was induced in hepatoma cell lines Huh7 and HepG2 cells, which suggested that Rock2 may be a new target for HCC gene therapy.

关 键 词：癌 肝细胞 细胞增殖 细胞凋亡 肝肿瘤 蛋白质丝氨酸苏氨酸激酶 转染 基因沉默 

分 类 号：R735.7[医药卫生—肿瘤]