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腺病毒载体携带人TRAIL基因治疗H446小细胞肺癌的体外实验研究
引用本文:刘永靖,陈飞虎,于奇,韩继彪,苏长青,钱其军.腺病毒载体携带人TRAIL基因治疗H446小细胞肺癌的体外实验研究[J].中国药理学通报,2007,23(6):755-759.
作者姓名:刘永靖  陈飞虎  于奇  韩继彪  苏长青  钱其军
作者单位:1. 解放军第105医院胸心外科,安徽,合肥,230031;安徽医科大学药学院,安徽,合肥,230032
2. 安徽医科大学药学院,安徽,合肥,230032
3. 解放军第105医院胸心外科,安徽,合肥,230031
4. 第二军医大学东方肝胆外科医院病毒基因实验室,上海,200438
摘    要:目的评价携带人TRAIL基因的腺病毒载体对人小细胞肺癌细胞株H446的基因治疗功效。方法构建的携带人TRAIL基因的腺病毒Ad-hTRAIL,感染人小细胞肺癌细胞株H446以及人正常肝细胞株WRL-68,人正常成纤维细胞株MRC-5,通过四甲基偶氮唑蓝(MTT)比色法检测其杀伤肿瘤细胞的效能,流式细胞术(FCM)检测Ad-hTRAIL对细胞早期凋亡的影响,并进行RT-PCR、酶联免疫吸附试验(ELISA)检测TRAIL mRNA和TRAIL蛋白的表达量。结果携带人TRAIL基因的增殖缺陷型腺病毒Ad-hTRAIL对H446细胞的感染效率较高;感染72h后H446、WRL-68、MRC-5细胞培养上清中TRAIL的表达量分别为32.02、17.08、16.89μg.L-1,在MOI=1.0时,Ad-hTRAIL即可引起H446细胞明显凋亡;而在MOI=100时对正常细胞WRL-68、MRC-5也没有明显杀伤作用。结论Ad-hTRAIL能够强烈诱导小细胞肺癌细胞H446凋亡而对正常肝细胞及成纤维细胞无明显抑制作用,Ad-hTRAIL在小细胞肺癌的基因治疗方面有潜在的应用前景。

关 键 词:小细胞肺癌  基因治疗  腺病毒  TRAIL  凋亡
文章编号:1001-1978(2007)06-0755-05
修稿时间:2007-01-01

Antitumor activity of replication-defective adenovirus carrying human tumor necrosis factor-related apoptosis-inducing ligand gene on human small cell lung cancer cell line H446 in vitro
LIU Yong-jing,CHEN Fei-hu,YU Qi,HAN Ji-biao,SU Chang-qing,QIAN Qi-jun.Antitumor activity of replication-defective adenovirus carrying human tumor necrosis factor-related apoptosis-inducing ligand gene on human small cell lung cancer cell line H446 in vitro[J].Chinese Pharmacological Bulletin,2007,23(6):755-759.
Authors:LIU Yong-jing  CHEN Fei-hu  YU Qi  HAN Ji-biao  SU Chang-qing  QIAN Qi-jun
Abstract:Aim To evaluate the therapeutic efficiency of Ad-hTRAIL (the replication-defective adenovirus expressing recombinant human TRAIL gene) on human small cell lung cancer (SCLC) cell line H446 in vitro. Methods Human SCLC cell line H446 and normal hepatocyte line WRL-68, fibroblast line MRC-5 were transfected with Ad-hTRAIL. The cytotoxicity in cultured SCLC and normal cells was evaluated by MTT. Flow Cytometry (FCM) was used to detect the early apoptotic induced by Ad-hTRAIL. RT-PCR, ELISA assay was used to detect the expression of TRAIL mRNA and protein. Results Ad-hTRAIL could be transfected into H446 cell line efficiently. Obvious apoptosis in cultured H446 cells was induced by Ad-hTRAIL at MOI of 1.0, but no significant effect on WRL-68 and MRC-5 even at MOI of 100. TRAIL expressed at high level in H446 cells but low in normal cells. 72 h after infected with Ad-hTRAIL, the expression of TRAIL in the supernatant of cultured cells H446, WRL-68 and MRC-5 were 32.02, 17.08 and 16.89 μg·L-1, respectively. Conclusion Ad-hTRAIL can induce apoptosis and growth suppression hardly on human SCLC cell line H446 but not on normal hepatocytes and fibroblasts. Replication-defective adenoviral vector carrying human TRAIL gene holds great promising in human SCLC therapy.
Keywords:TRAIL
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