摘要
目的:观察人突变p27基因(p27mt)对人大肠癌细胞生长的影响,探讨p27mt基因在大肠癌基因治疗中的作用机制。方法:以携带突变p27基因复制缺陷型腺病毒(Ad-p27mt)为载体,转染大肠癌细胞SW480;用Westernblot方法检测p27mt蛋白的表达;细胞计数法检测p27mt对SW480细胞生长的抑制作用;用流式细胞仪检测细胞周期;用DNA片段分析法检测细胞凋亡。结果:通过免疫印迹分析Ad-p27mt转染SW480细胞后,p27在细胞中出现了蛋白高表达。PI染色流式细胞仪检测77.96%的细胞阻滞于G0/G1期,而Ad-LacZ组及空白对照组分别为27.57%和25.29%;生长曲线显示Ad-p27mt对细胞生长就有明显的抑制作用;DNA片段分析示p27mt基因可诱导细胞的凋亡。结论:p27mt对细胞周期有明显的阻滞作用,主要阻滞于G0/G1期;p27mt基因对细胞的生长抑制机制与诱导细胞凋亡和细胞周期的阻滞有关。
Objective： To study the effect of the mutant p27 gene （p27mt gene） on growth of the colorectal cancer cell line SW480 and to evaluate the therapy significance by DNA ploidy analysis and apoptosis measuration. Methods： Adenovirns vector with exogenous mutant p27 gene was transfected into the colorectal cancer cell line SW480, the transfer efficiency and expression effect of p27mt gene were examined by X-gal stains and western blot analysis, the growth of SW480 in vitro was determined by cell counting. Flow cytometric analysis was used for observing cell cycle. The Apoptosis was measured by DAN fragment analysis. Results： Transfer efficiency reached 100% in SW480 when adenovirns infection ability was over 50MOI, the efficiency expression of p27mt gene was measured using western blot. A 77.96% of cells was arrested in G0/G1 in experimental group,while Ad-LacZ group and blank contrast group were 27.57% and 25.29% in G0/G1. respectively. The curve of growth showed that p27mt gene had a inhibition effect on SW480. DNA fragment analysis demonstrated that p27mt could induce SW480 cell apoptosis. Conclusion： Adenovirns gene transfer of p27mt induces the growth suppression and the cell cycle arrest in G1-S and apoptosis of colorectal cancer cell line SW480. The mechanism of growth suppression may be related to apoptosis and cell cycle arrest.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2005年第15期849-851,共3页
Chinese Journal of Clinical Oncology
基金
湖北省自然科学基金资助(编号:2003ABA193)
关键词
大肠癌
p27mt基因
生长抑制
凋亡
Colorectal cancer p27 mt gene Growth suppression Apoptosis
