Mus81基因沉默对Hep3B人肝癌细胞系增殖和凋亡的影响  

Effects of methyl methansulfonate and UV sensitive gene clone 81 on proliferation and apoptosis of human hepatocellular carcinoma Hep3B cells

作　　者：张芳雍 谭国钳 曾裕文 吴帆[1] ZHANG Fangyong;TAN Guoqian;ZENG Yuwen;WU Fan(Department of Hepatobiliary Surgery,Guangzhou Red Cross Hospital,Ji′nan University,Guangzhou 510220,China)

机构地区：[1]暨南大学医学院附属广州红十字会医院肝胆外科,广州510220

出　　处：《实用医学杂志》2020年第23期3189-3193,共5页The Journal of Practical Medicine

基　　金：国家自然科学基金面上项目(编号:81974442);广东省自然科学基金面上项目(编号:2020A1515010799);广州市科学研究计划项目(编号:201804010025)。

摘　　要：目的研究甲磺酸盐及紫外线敏感性81号基因(methyl methanesulfonate and UV sensitive gene clone 81,Mus81)沉默对Hep3B人肝癌细胞系增殖和凋亡的影响。方法采用慢病毒介导的小干扰RNA技术沉默Mus81基因的表达并以实时荧光定量聚合酶链反应(qRT⁃PCR)检测Mus81基因的沉默效率。噻唑蓝(MTT)法绘制细胞生长曲线,平板克隆实验、碘化丙锭(PI)染色法和膜联蛋白V(Annexin V)⁃APC染色法分别检测Mus81沉默对Hep3B细胞系增殖、细胞周期及凋亡的影响。结果慢病毒感染效率高于80%,Mus81基因干扰效率为76.42%(P<0.001)。MTT生长曲线和平板克隆实验的结果显示,Mus81沉默的Hep3B细胞系的生长速度明显下降(P<0.001),增殖能力也明显减弱[细胞克隆数:(2±1)比(54±3),P<0.001]。Annexin V⁃APC和PI染色检测显示Mus81沉默Hep3B细胞系的凋亡率明显升高[(20.87±0.82)%vs.(0.69±0.09)%,P<0.001]并出现G1期细胞比例轻微降低[(33.77±1.61)%vs.(42.06±0.40)%,P<0.05]。结论Mus81基因沉默可明显抑制人肝癌细胞系Hep3B的生长增殖并促进其凋亡。Objective To study the effects of methyl methanesulfonate and UV sensitive gene clone 81(Mus81)silencing on proliferation and apoptosis of human hepatocellular carcinoma Hep3B cells.Methods Lenti⁃virus⁃mediated small interfering RNA(siRNA)was employed to inhibit Mus81 expression and real⁃time quantitative polymerase chain reaction(Real⁃time PCR)was used to determine the inhibition efficiency of Mus81 expression.Growth curve was established by the methylthiazol tetrazolium(MTT)assay.The plate cloning experiment,propidium iodide(PI)and Annexin V⁃APC staining were used to detect the proliferation,cell cycle distribution and apoptosis of Hep3B cell line.Results The efficiency of lentivirus infection was higher than 80%,and the efficiency of Mus81 gene interference was 76.42%(P<0.001).The results of MTT growth curve and plate cloning experiment showed that the growth rate of the Hep3B cell line silenced by Mus81 was decreased significantly(P<0.001),the proliferation ability was also significantly weakened(number of cell clones:(2±1)vs.(54±3),P<0.001).Annexin V⁃APC and PI staining showed that the apoptosis rate of Mus81 silenced Hep3B cell line was significantly increased[(20.87±0.82)%to(0.69±0.09)%,P<0.001]and the proportion of cells in G1 phase was slightly decreased[(33.77±1.61)%to(42.06±0.40)%,P<0.05].Conclusion Mus81 gene silencing can significantly inhibit the growth and proliferation of human liver cancer Hep3B cells and promote its apoptosis.

关 键 词：甲磺酸盐及紫外线敏感性81号基因 肝细胞癌 增殖 凋亡 小干扰RNA 

分 类 号：R735.7[医药卫生—肿瘤]