摘要
目的 探讨p2 1waf1基因过表达在人胆管癌细胞增殖和凋亡中的作用。方法 包含 2 .1kb的 p2 1waf1cDNA片段经阳离子脂质体DOTAP包裹转染人胆管癌细胞QBC939中 ,并得到稳定表达 ,通过聚合酶链反应 (PCR)、逆转录 PCR(RT PCR)、流式细胞仪及免疫组织化学等方法检测p2 1waf1基因表达、细胞生长和细胞凋亡等情况。结果 p2 1waf1基因转染后 ,细胞生长明显受抑制 ,细胞周期停滞于G1期 ,表现为与对照组相比 ,细胞周期G1期比例明显增加 ,S和G2 /M期比例明显减少(G1期 :5 7.32 %→ 73 .6 9% ;S期 :2 1.81%→ 15 .85 % ;G2 /M期 :19.35 %→ 11.76 % ) ,细胞凋亡率增加(3 .47%→ 15 .81% ) ,裸鼠致瘤性降低。结论 p2 1waf1基因能够抑制人胆管癌细胞增殖和诱导细胞凋亡。
Objective\ To explore the role of p21 waf1 overpression in the proliferation and apoptosis of human cholangiocarcinoma. Methods\ p21 waf1 gene packaged with lipofectin DOTAP was transfected into the cells of a human cholangiocarcinoma cell line (QBC939), which stably expressed p21 waf1 gene. The p21 waf1 mRNA and protein expression of QBC939 p21 waf1 , QBC939 Vect and QBC939 cell were detected by RT PCR and immunocytochemical methods, respectively. The cell cycle pattern and apoptosis were assayed by using flow cytometry. Results\ After transfection of p21 waf1 , cell growth was obviously inhibited and resulted in an accumulation of cells in G 1, presenting that the proportion of the cells in G 1 phase was obviously increased from 57.32 % to 73.69 %, and that of the cells in S and G 2/M phases was obviously decreased from 21.81 % to 15.85 % and from 19.35 % to 11.76%, respectively, as compared with control group. The apoptotic index was increased in QBC939 p21 waf1 cells from 3.47 % to 15.81 %. Tumorigenicity of the QBC939 p21 waf1 cells in nude mice was greatly inhibited. Conclusion\ Tranfer of p21 waf1 gene could inhibit the growth of cholangiocarcinoma cells and induce cell apoptosis.\;
出处
《中华实验外科杂志》
CSCD
北大核心
2000年第6期514-517,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金!资助项目 (39770 72 7)
