Bad联合Caspase-8促凋亡基因共表达对SK-HEP-1肝癌细胞体外增殖、凋亡、迁移和体内成瘤影响的实验研究  

The experiment of Bad and Caspase-8 gene Co-expression on proliferation,apoptosis,migration and tumorigenesis of SK-HEP-1 hepatoma carcinoma cells

作　　者：鲁斌[1] 程敏[1] 周凡[2] LU Bin;CHENG Min;ZHOU Fan(Department of Hepatobiliary Surgery,Chaohu Hospital Affiliated to Anhui Medical University,Chaohu 238000,China;不详)

机构地区：[1]安徽医科大学附属巢湖医院肝胆外科,安徽巢湖238000 [2]南昌大学第二附属医院,南昌330006

出　　处：《实用医学杂志》2020年第8期1030-1034,共5页The Journal of Practical Medicine

基　　金：江西省科技计划项目(编号:20151BBG70042)。

摘　　要：目的观察Bad联合Caspase-8促凋亡基因共表达对SK-HEP-1肝癌细胞体外增殖、凋亡、迁移和体内成瘤影响。方法利用基因重组技术构建Bad-Caspase-8共表达融合基因。慢病毒转染SK-HEP-1肝癌细胞72 h后,蛋白质免疫印迹法观察细胞中Bad蛋白和Caspase-8蛋白表达,CCK-8法检测BadCaspase-8融合基因过表达对SK-HEP-1肝癌细胞增殖影响,流式细胞仪检测其对SK-HEP-1肝癌细胞凋亡影响,Transwell侵袭实验检测转染后细胞迁移变化,最后将转染SK-HEP-1肝癌细胞接种至裸鼠皮下观察成瘤效果。结果转染72 h后,SK-HEP-1肝癌细胞中Bad蛋白和Caspase-8蛋白水平显著增加,细胞增殖能力显著降低,实验组、对照组和空白组OD值分别为0.86±0.122、1.89±0.315和2.01±0.194,P<0.001;转染后SK-HEP-1肝癌细胞凋亡率明显增加,实验组、对照组和空白组细胞凋亡率分别为(31.6±2.8)%、(3.2±0.9)%和(2.9±1.1)%,P<0.001,转染后SK-HEP-1肝癌细胞迁移能力显著降低,实验组、对照组和空白组细胞迁移数分别为18±3,77±10和85±11,P<0.001;细胞接种至裸鼠皮下15 d,与对照组和空白组相比,实验组肿瘤生长速度缓慢,实验组、对照组和空白组成瘤体积分别为(1.6±0.17)cm3、(4.4±0.29)cm3和(4.9±0.42)cm3,P<0.001。结论促凋亡基因Bad和Caspase-8共表达可显著降低SK-HEP-1肝癌细胞增殖活性,促进SK-HEP-1肝癌细胞凋亡,减少SK-HEP-1肝癌细胞迁移,抑制SK-HEP-1肝癌细胞体内成瘤效率,可作为肝癌靶向治疗潜在靶点。ObjectiveTo explore the effect of Bad and Caspase-8 gene Co-expression on the proliferation,apoptosis,migration and tumorigenesis of SK-HEP-1 hepatoma carcinoma cells in vitro and in vivo.Methods Building Bad-Caspase-8 fusion gene using gene recombination technology.72 hours after SK-HEP-1 hepatoma carcinoma cells transfection,the expression of Bad and Caspase-8 protein was measured with Western Blot,the cell proliferation was detected by CCK-8,the cell apoptosis was tested by flowcytometry,the cell migration was detected by Transwell invasion,and finally,the transfected SK-HEP-1 hepatocellular carcinoma cells were inoculated under the skin of nude mice to observe the tumorigenesis in vivo.Results Western Blot showed that the expression of Bad and Caspase-8 protein was increased significantly in SK-HEP-1 hepatocellular carcinoma cells at72 h after transfection.Compared with the control group and the blank group,the cell proliferation ability of the experimental group was significantly decreased at 72 hours after transfection,the OD value of experimental group,control group and blank group were 0.86±0.122,1.89±0.315 and 2.01±0.194,P<0.001.The cell apoptosis rate in the experimental group was obviously higher than other groups,the cell apoptosis rate of experimental group,control group and blank group were(31.6±2.8)%,(3.2±0.9)%and(2.9±1.1)%,P<0.001.The migration ability of SK-HEP-1 hepatocellular carcinoma cells was significantly reduced,the number of cell migration of experimental group,control group and blank group were 18±3,77±10 and 85±11,respectively,P<0.001.The tumor growth rate of the experimental group was slower than that of the control group and the blank group.The tumor volume of experimental group,control group and the blank group were(1.6±0.17)cm3,(4.4±0.29)cm3 and(4.9±0.42)cm3,respectively,P<0.001.ConclusionsBad and Caspase-8 gene Co-expression can inhibit SKHEP-1 hepatocellular carcinoma cell proliferation,induce SK-HEP-1 hepatocellular carcinoma cell apoptosis and reduce SK-HEP-1 hepatoc

关 键 词：BAD CASPASE-8 细胞凋亡 细胞增殖 细胞迁移 移植成瘤 

分 类 号：R735.7[医药卫生—肿瘤]