去甲斑蝥素对人类胆管癌细胞系QBC939增殖和凋亡作用的实验研究

 【摘要】　目的　研究去甲斑蝥素（NCTD）对人类胆管癌细胞系QBC939的生长抑制作用，初步探讨其作用机制。方法　将NCTD作用于经体外培养的QBC939细胞系，分别采用四甲基偶氮唑蓝比色法（MTT法），流式细胞术及免疫组织化学SP法进行检测。结果　0.125、0.75、2.5、10、120 μg／ml NCTD作用48 h，对QBC939细胞系均有抑制作用（P＜0.05），且具有浓度依赖性，绘制浓度效应曲线，得出半数抑制浓度（IC50）为（3.66±1.14）μg／ml；在分别作用12、24、36、48、72 h后，生存率有随时间增加而下降的趋势（P＜0.05）。流式细胞术结果表明，随着NCTD浓度的增加，凋亡率逐渐增高，各浓度组分别为（8.6±0.4）%、（17.6±0.3）%、（22.9±0.4）%、（25.5±0.9）%、（31.1±1.5）%（P＜0.001）；质量浓度为2.5 μg／ml的NCTD作用QBC939细胞48 h后，出现G2／M期阻滞现象，NCTD与对照组分别为（14.1±1.0）%和（5.7±0.3）%（P＜0.05）。不同浓度的NCTD作用于QBC939细胞后，与对照组相比Caspase-3蛋白表达均增加。结论　NCTD具有抑制人类胆管癌细胞系QBC939增殖的作用，这种作用可能与诱导细胞凋亡，干扰细胞生长周期有关。

Experimental study of norcantharidins effects on proliferative growth and apoptosis of human cholangiocarcinoma cell line QBC939

Objective To study the growth effect of human cholangiocarcinoma cell line QBC939 treated by norcantharidin (NCTD) and preliminary illustrate the potential mechanism.Methods The human cholangiocarcinoma cell line QBC939 was detected by MTT assay,flow cytometry,immunocytochemistry after the treatment of NCTD in vitro.Results NCTD displayed inhibitory effect on growth of QBC939 from different doses of 0.125,0.75,2.5,10,120 μg/ml after 48 h (P ＜0.05).It was in a dose and time dependent manner.Dose-effect curve was drawn and IC50 value was (3.66±1.14) μg/ml.The flow cytometric profiles showed that the rate of cell apoptosis enhanced following increasing the concentration of NCTD(8.6±0.4) ％,(17.6±0.3) ％,(22.9±0.4) ％,(25.5±0.9) ％ and (31.1±1.5) ％,respectively]and cells blocked in the G2/M phase after treatment with 2.5 μg/ml NCTD(14.1±1.0) ％ and (5.7±0.3) ％].The expression of the protein caspase-3 elevated after different concentrations of NCTD co-cultured with QBC939 compare with contrast group.Conclusion NCTD has an inhibitory effect on proliferation of QBC939 cell line,and the mechanism might be related to the induction of cell apoptosis and blockade of cell cycle.