人突变p27基因对大肠癌细胞凋亡的影响

Effects of Mutant p27 Gene on the Apoptosis of Colorectal Cancer Cell

目的探讨人突变p27基因对大肠癌细胞凋亡的诱导作用。方法构建重组腺病毒Ad-p27mt,转染大肠癌细胞SW480,用流式细胞仪、DNA片段分析法、TUNEL法观察Ad-p27mt对大肠癌细胞的凋亡的诱导作用。结果成功构建了Ad-p27mt,在感染复数≥50时,可达100%的转导效率。Ad-p27mt转染SW480。24h后流式细胞仪检测在G1期前出现了亚二倍体凋亡峰,细胞DNA提取电泳后出现了凋亡特征性梯带,TUNEL法检测凋亡指数实验组和对照组分别为(82.0±3.2)和(5.0±3.5),差异有显著性(P<0.01)。结论人突变p27基因能有效诱导大肠癌细胞的凋亡。

Objective To study the inducing effect of human mutant p27 gene on apoptosis of the colorectal cancer cells. Methods The recombinant Adenovirus Ad-p27mt was constructed to infect the colorectal cancer cell SW480. The inducing effect of Ad-p27 mt on apoptosis of the colorectal cancer was measured by the flow cytometer, the DNA fragment analysis and TUNEL method. Results Ad-p27mt was successfully constructed. If the multiplicity of infection （MOI） was ≥50, the infection efficiency can reach 100%. After 24h of infection, there was an apoptotic hypodiploid peak on the flow cytometer before G1 and there were apoptotic characteristic bands in the DNA electrophoresis. The apoptotic index detected by TUNEL method was 82. 6 ± 3. 2 （Ad-p27mt group） and 5. 0 ± 3. 5 （control group） respectively, the difference of which was significant （P〈0. 01）. Conclusion Human mutant p27 gene can effectively induce apoptosis of the colorectal cancer cells.