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首页> 美国卫生研究院文献>World Journal of Gastroenterology >Tumor necrosis factor-related apoptosis-inducing ligand gene on human colorectal cancer cell line HT29
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Tumor necrosis factor-related apoptosis-inducing ligand gene on human colorectal cancer cell line HT29

机译：大肠癌细胞HT29的肿瘤坏死因子相关的凋亡诱导配体基因

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AIM: To evaluate the therapeutic efficiency of Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) gene on human colorectal cancer cell line HT29.METHODS: Human embryonal kidney cells transformed by introducing sheared fragments of Ad5 DNA (293 cell) were used for amplification of adenoviral vectors: Ad/GT-TRAIL, Ad/GT-Bax, Ad/GT-LacZ and Ad/PGK-GV16. Human colorectal cancer cell line HT29 was transfected with binary adenovirus-mediated TRAIL gene. Bax gene was used as positive control, LacZ gene was used as the vector control, and cells treated with PBS only were used as a mock control. The morphological changes, cell growth and apoptosis were measured by reversmicroscope, MTT method and flow cytometry.RESULTS: All adenoviral vectors titer determined by optical absorbency at A260nm were 1 × 1010 viral particle/ml(vp/ml). Obviously morphological changes of HT29 cells were observed when infected with Ad/GT-TRAIL, and these changes were much more obviously when Ad/PGK-GV16 was coinfected. The cell suppression percentage and the percentage of apoptotic cells were 52.5% and 16.5% respectively when infected with Ad/GT-TRAIL alone, while combining with Ad/PGK-GV16, the growth of HT29 was suppressed by 85.2% and the percentage of apoptotic cells was 35.9%. It showed a significantly enhanced therapeutic efficiency with binary system (P < 0.05).CONCLUSION: A binary adenoviral vector system provides an effective approach to amplify viral vectors that express potentially toxic gene, TRAIL. Ad/GT-TRAIL showed a significantly enhanced therapeutic efficiency for HT29 when coinfected with Ad/PGK-GV16. Ad/GT-TRAIL could induce apoptosis of HT29 and inhibit its growth.
机译：目的：评价肿瘤坏死因子相关的凋亡诱导配体（TRAIL）基因对人结肠直肠癌HT29细胞的治疗效果。方法：采用经剪切的Ad5 DNA片段（293细胞）转化的人胚肾细胞。腺病毒载体的扩增：Ad / GT-TRAIL，Ad / GT-Bax，Ad / GT-LacZ和Ad / PGK-GV16。用二元腺病毒介导的TRAIL基因转染人结肠直肠癌细胞系HT29。 Bax基因用作阳性对照，LacZ基因用作载体对照，仅用PBS处理的细胞用作模拟对照。结果：通过A260nm吸光度测定的所有腺病毒载体滴度均为1×10 10 病毒颗粒/ ml（vp /毫升）。感染Ad / GT-TRAIL时，可以观察到HT29细胞的形态学变化，而同时感染Ad / PGK-GV16时，这些变化更明显。单独感染Ad / GT-TRAIL时，细胞抑制率和凋亡细胞率分别为52.5％和16.5％，而与Ad / PGK-GV16结合时，HT29的生长被抑制85.2％，凋亡率被抑制细胞为35.9％。结论：二元腺病毒载体系统为扩增表达潜在毒性基因TRAIL的病毒载体提供了有效的方法。当与Ad / PGK-GV16共感染时，Ad / GT-TRAIL对HT29的治疗效率显着提高。 Ad / GT-TRAIL可诱导HT29凋亡并抑制其生长。

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