腺病毒介导的分化基因RA-538诱导肝癌细胞增殖抑制和凋亡的研究  

Study on the induction of proliferative inhibition and apoptosis of hepatocellular carcinoma cells by adenovirus mediated differentiation gene RA538

作　　者：张海增[1] 林晨[2] 邵永孚[1] 隗月[2] 付明[2] 张雪艳[2] 梁肖[2] 吴旻[2] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤医院腹部外科,北京100021 [2]中国医学科学院中国协和医科大学肿瘤医院分子肿瘤学国家重点实验室,北京100021

出　　处：《中华实验外科杂志》2001年第5期453-455,共3页Chinese Journal of Experimental Surgery

基　　金：国家 8 6 3高科技发展基金资助项目 (Z 2 0 0 1 0 2 )

摘　　要：目的　研究携带分化基因RA 53 8的重组腺病毒 (Ad RA53 8)对人肝癌细胞的抑制作用及机制。方法　Ad RA53 8感染人肝癌细胞系BEL 740 2和QSG 770 1 ,绘制生长曲线 ,克隆形成实验评价Ad RA53 8对肝癌细胞增殖的抑制作用 ,应用流式细胞术、DNA原位末端标记、梯形DNA研究细胞周期及细胞凋亡情况 ,逆转录 聚合酶链式反应 (RT PCR)和免疫印迹法 (Westernblot)检测相关基因的表达。结果　感染Ad RA53 8后 ,RA53 8在肝癌细胞内获得了表达 ,并且降低了c myc的表达。肿瘤细胞的增殖受到明显抑制 ,感染Ad RA53 85d后 ,BEL 740 2的活细胞数为(1 0 .0± 3 .7)× 1 0 4,对照组为 (1 72 .7± 1 1 .9)× 1 0 4(P <0 .0 0 1 ) ;QSG 770 1感染Ad RA53 83d后的活细胞数为 (2 3 .0± 4.3 )× 1 0 4,对照组为 (91 .7± 7.2 )× 1 0 4(P <0 .0 0 1 ) ;感染Ad RA53 8后BEL 740 2和QSG 770 1形成的克隆数分别为 1 3± 4和 1 5± 2 ,明显少于对照组的 1 88± 9和 1 75± 9(P <0 .0 0 1 ) ,而且还出现了细胞周期的阻滞和凋亡。结论　Ad RA53 8对肝癌细胞具有明显的抑制作用 。Objective To investigate the inhibitory effect on human hepatocellular carcinoma cells and the mechanism of the recombinant adenovirus encoding differentiation gene RA538 (Ad RA538).Methods Human hepatocellular cancer cell lines BEL 7402 and QSG 7701 were infected with Ad RA538. Cell growth curve and cloning formation test was used to investigate the inhibitory effects of Ad AR538 on the growth of hepatocellular cancer cells. Flow cytometry, DNA ladder and tunnel assay were used to study the cell cycle and apoptosis. RT PCR and Western blot were used to detect the related gene expression.Results After infection of Ad RA538, the high expression of RA 538 and decreasing expression of c myc was observed in the hepatocellular cancer cell lines.Ad RA538 could obviously inhibit the growth of the two hepatocellular cancer cells. Five days after transfected with Ad RA538, the cell number of BEL 7402 was (10.0±3.7)×10 4, while (172.7±11.9)×10 4 (P<0.001) in the control group; Three days after transfected with Ad RA538, the cell number of QSG 7701 was (23.0±4.3)×10 4, while (91.7±7.2)×10 4 in the control group (P<0.001). The cloning number of BEL 7402 and QSG 7701 transfected with Ad RA538 was 13±4 and 15±2 respectively, but the cloning number of them transfected with Ad LacZ was 188±9 and 175±9 (P<0.001) respectively. Furthermore, Ad RA538 could induce cancer cell cycle arrest and terminal differentiation (apoptosis).Conclusion Ad RA538 has obviously inhibitory effects on hepatocellular cancer cell and has great potential value in clinical cancer therapy.

关 键 词：重组腺病毒载体 基因治疗 RA538基因 肝癌细胞 

分 类 号：R735.7[医药卫生—肿瘤]