摘要
目的 探讨nm23-H1基因转染对人胆管癌细胞系增殖能力的影响。方法 通过脂质体法将含有全长nm23-H1 cDNA的真核表达载体转染人胆管癌细胞系QBC939,Northern blotring及免疫组织化学观察转染前后细胞内nm23-H1基因的mRNA及蛋白表达情况,噻唑蓝(MTT)比色法检测转染前后细胞体外增殖能力变化,流式细胞仪分析转染前后细胞周期及凋亡改变。结果 转染成功的QBC_(939)-nm23细胞nm23-H1基因的mRNA及蛋白表达明显上升,体外增殖能力下降,G_0/G_1期、G_2/M期、S期比例及凋亡率(％)分别为(70.24±2.05、12.37±0.91、17.39±2.95、9.54±0.78),与亲本QBC_(939)细胞(56.57±0.99、16.00±1.39、27.44±2.09、4.23±1.05)及对照组QBC_(939)-C细胞(56.17±0.82、17.23±0.77、26.60±0.53、3.67±0.49)相比,G_0/G_1期细胞比例明显增加,G_2/M期及S期细胞比例明显下降,凋亡细胞增加(P<0.05)。结论 nm23-H1基因可以抑制体外胆管癌细胞的增殖能力及诱导凋亡。
Objective To explore the role of nm23-H1 gene overexpression in proliferation of human cholangiocarcinoma cell line. Methods A mammalian expression vector containing nm23-H1 cDNA was transfected into human QBC_(939) cholangiocarcinoma cell line. Tests such as Northern blotting and immunohistochemistry were used to analyze the expression levels of nm23-H1. Flow cytometry and MTT were used to analyze the changes of cell cycle, apoptosis and proliferation of the tumor cells before and after the gene transfection. Results After transfection, the nm23-H1 mRNA and protein expression were upregulated significantly. The ability of proliferation was remarkably decreased. The percent of G_0/G_1, G_2/M,S period and apoptosis was (70.24 ± 2.05)%, (12.37 ± 0.91)%, (17.39 ± 2.95)% and (9.54 ± 0.78)% respectively. Compared with QBC_(939) cell and QBC_(939)-C cell, the percent of G_2/M and S period were decreased and the percent of G_0/G_1 period was increased. The percent of apoptosis was increased. Conclusion Up-regulation of nm23-H1 could suppress the abilities of proliferation and induce apoptosis in cholangiocarcinoma cell line in vitro.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2004年第8期948-950,共3页
Chinese Journal of Experimental Surgery
基金
全军医药卫生科研基金(96Q067)
重庆市科委应用基础研究项目(CQ19980067)
