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及可能机制。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;Western blotting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:Ad-p27mt转染SW480细胞后,p27在细胞中出现了蛋白高表达;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。
AIM： To observe the influence of human mutant p27 gene （p27mt） on the growth and so as to investigate the function and mechanism of p27mt in gene therapy for colorectal cancer. METHODS： Colorectal cancer cell line SW480 was infected with recombinant replication defective adenovirns Ad - p27mt, and expression of p27mt protein was detected by Western blotting. The inhibitory effect of p27mt on SW480 and cell cycle were determined by flow cytometry, and DNA fragment was analyzed to identify the occurrence of apoptosis. RESULTS： After transfected with Ad - p27mt, p27 protein was highly expressed in SW480 cells, 77, 96% colorectal cancer cells were blocked in phase G0/G1 , while in Ad - LacZ group and blank control group, 27.57% and 25.29% cells were blocked in the same phase, respectively, Growth curve showed Ad - p27mt had an obviously inhibitory effect on the growth of SW480 cells, DNA fragment assay demonstrated that p27mt was able to induce the apoptosis of colorectal cancer cells, CONCLUSION： p27mt has an obvious blocking effect on colorectal cancer cell cycle, and most cells are blocked in phase G0/G1. This blockage is related with the growth inhibition and apoptosis induced by p27mt,
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第10期2039-2041,共3页
Chinese Journal of Pathophysiology
基金
湖北省自然科学基金资助项目(No.2003ABA193)
