人增殖抑制基因(hHSG)联合热处理对结肠癌细胞系HT-29的体外诱导凋亡效应  

Transfection of human hyperplasia suppressor gene and hyperthermia have a synergistically apoptotic effect on colon cancer cell HT-29 in vitro

作　　者：毋丽娜[1] 李志新[1] 卢恬[1] 马腾[1] 邱晓彦[1] 

机构地区：[1]北京大学基础医学院免疫学系

出　　处：《北京大学学报（医学版）》2008年第5期524-527,共4页Journal of Peking University:Health Sciences

基　　金：国家高技术研究发展计划专项经费(2002AA216131)资助~~

摘　　要：目的:观察人增殖抑制基因(human hyperplasia suppressor gene,hHSG)超表达联合热处理对结肠癌细胞株HT-29的细胞存活及其诱导凋亡的效应。方法:首先制备复制缺陷5型腺病毒-hHSG表达载体(Ad5-hHSG),然后将其在HT-29中超表达,观察Ad5-hHSG对肿瘤细胞的热增敏效应。采用细胞计数法检测其对细胞存活的影响,流式细胞术检测细胞凋亡和周期改变。结果:100感染复数(multiplicity of infection,MOI)剂量Ad5-hHSG与42℃加热1h联合应用后,活细胞数显著低于单用加热组和单用Ad5-hHSG组(P<0.01)。流式细胞术检测细胞凋亡,Ad5-hHSG单独作用组、单独加热组以及联合作用组细胞凋亡率均较对照组显著升高,而且联合作用组与单独作用组之间差异具有统计学意义(P<0.01)。细胞周期结果显示,Ad5-hHSG与42℃加热1h联合作用可明显导致HT-29细胞阻滞在G2/M期。结论:Ad5-hHSG对肿瘤细胞具有热增敏效应。Objective：To investigate the suppression effect and apoptosis of the colon cell line HT-29 treated with adenovirus-expressed human hyperplasia suppressor gene （hHSG） and hyperthermia. Methods：Replication-defective recombinant adenovirus containing hHSG was constructed, and then infected into HT-29 cell line to detect its overexpression by western blot. The synergic effect with hyperthermia was observed, cell survival was assessed using Vi-CELL TMXR Cell Viability Analyzer, and cell apoptosis and cell cycle distribution were determined by flow cytometry. Results： After the cells were treated with 100 MOI （multiplicity of infection） Ad5-hHSG and heated at 42℃ for 1 h, the number of viable ceils decreased sharply, compared with single treatment with AdS-hHSG or 42℃ for 1h （P〈0.01）. The apoptotic rate of combined groups was significantly higher than that of Ad5-hHSG group or the heated group respectively （P〈0.01）. Moreover, the combination treatment of Ad5-hHSG and 42℃ for 1h induced G2/M arrest. Conclusion： Ad5-hHSG could increase the sensitivity of HT-29 to hyperthermia.

关 键 词：基因 抑制 高温 诱发 细胞凋亡 细胞增殖 

分 类 号：R73-351[医药卫生—肿瘤]