凋亡素基因质粒载体对人肝癌细胞的影响  

The effect of expression of apoptin gene on human hepatoma cell in vitro

作　　者：黄勋[1] 蔡筱彦[1] 顾红祥[1] 王立生[1] 朱惠明[1] 

机构地区：[1]暨南大学医学院附属深圳市人民医院消化科,518020

出　　处：《现代消化及介入诊疗》2003年第2期66-68,共3页Modern Interventional Diagnosis and Treatment in Gastroenterology

摘　　要：目的探讨凋亡素基因表达对人肝癌细胞的影响。方法将凋亡素基因克隆入真核表达载体pEGFP-C2,构建成pEGFP-VP3 为了使EGFP和EGFP-VP3在人肝癌细胞中表达,用非脂质体脂质转染法将pEGFP-C2和pEGFP-VP3分别转染人类肝癌细胞系(HepG2),通过荧光显微镜观察EGFP和EGFP-VP3在HepG2中的定位、表达、细胞生长及凋亡,用Annexin V-藻红素(PE)检测细胞凋亡。结果在HepG2/EGFP组细胞中,EGFP均匀分布于细胞浆和细胞核,细胞形态无变化。与非转染细胞的自然凋亡相比,转染细胞中细胞凋亡没有增加。在HepG2/EGFP-VP3组细胞中,EGFP-VP3以荧光颗粒形式集中在细胞核,细胞核中EGFP-VP3颗粒逐渐变粗,细胞变得小而圆,最后成为碎片,Annexin V-PE染色显示细胞凋亡;HepG2/EGFP-VP3组的细胞凋亡率明显高于HepG2/EGFP组和HepG2组(P<0.01)。结论 pEGFP-VP3转染人肝癌细胞后,凋亡素基因表达并导致癌细胞凋亡。Objective To investigate the effect of expression of apoptin gene on human hepatoma cells. Methods pEGFP-VP3 was constructed by cloning an apoptin gene to eukaryotic expression vector pEGFP-C2. In order to obtain expression of EGFP and EGFP-VP3 in human hepatocellular carcinoma cells, two plasmids, pEGFP-C2 and pEGFP-VP3, were transient transfected into HepG2, a human hepatocellular carcinoma cell line, by a non-liposome lipid transfection method, respectively. The location and expression of EGFP and EGFP-VP3 in HepG2, cell growth and apoptosis were observed. Apoptosis in HepG2 was assayed by Annexin V-PE dying. Results In HepG2/EGFP cells, EGFP was fairly well-distributed in both plasma and nucleus. And no change was found in cell morphology. The rate of apoptosis in EGFP in transfected cells was not increased as compared with that in non-transfected cells HepaG2. In HepG2/EGFP-VP3 cells, EGFP-VP3 was concentrated in nucleus as fluorescent granules. The EGFP-VP3 granules in the nucleus became coarse and the cells became small and round, and finally into pieces. Apoptosis in the cells of HepG2/EGFP-VP3 was shown by Annexin V-PE dying. Rate of apoptosis in HepG2/EGFP-VP3 cells was higher than those in both HepG2/EGFP and HepG2 (P<0.01). Conclusion Transient transfection and expression of apoptin gene in human hepatoma cells can induce apoptosis of the cells.

关 键 词：凋亡素基因 质粒载体 肝细胞癌 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]