蟾蜍灵对人胆管癌细胞QBC939增殖及cyclin E和P27蛋白表达影响的研究

Effects of bufalin on the proliferation and expressions of cyclin E and P27 in changliocarcinoma cell line QBC939

目的已有诸多研究显示中药蟾蜍灵具有明显抑制肿瘤增殖的作用,观察蟾蜍灵对人胆管癌细胞QBC939的cyclin E和P27表达的影响,探讨其调控胆管癌细胞增殖的途径和机制。方法 MTT比色法观察不同浓度蟾蜍灵(0.1、1、10μmol/L)对人胆管癌细胞QBC939增殖的影响;乳酸脱氢酶(lactate dehydrogenase,LDH)实验检测蟾蜍灵毒性;流式细胞术检测人胆管癌细胞QBC939细胞周期的变化;Western blot法测定cyclin E、P27蛋白水平变化。结果 MTT比色法结果表明蟾蜍灵对人胆管癌细胞QBC939生长具有较强的抑制作用,并在一定范围内具有时间和浓度依赖性,流式细胞仪检测细胞周期阻滞在G0/G1期,具有良好的量效关系。Western blot测定cyclin E蛋白表达量下降,P27蛋白表达量升高。结论蟾蜍灵可能是通过阻滞细胞周期,减少cyclinE蛋白表达,增加P27蛋白表达,来达到抗肿瘤作用。

Objective Many studies have shown that bufalin,as a Chinese medicine,can significantly inhibit the proliferation of various of tumor cells.This study was to observe the effects of bufalin on the expressions of p27 and cyclin E in Changliocarcinoma cell line QBC939,and to investigate the pathways of bufalin in inhibiting tumor cell proliferation and its antitumor mechanism.Methods Changliocarcinoma cell line QBC939 was cultured by the routine method,and then treated with bufalin at the concentrations of 0.1,1 and 10 μmol/L.Bufalin toxicity was detected by lactate dehydrogenase（LDH）,the proliferation,apoptosis and cell cycle of QBC939 cells investigated by MTT and flow cytometry,and the protein expressions of p27 and cyclin E were determined by Western.Results Bufalin remarkably inhibited the proliferation of Changliocarcinoma cell line QBC939 in a time-and dose-dependent manner.Flow cytometry showed the cell cycle was arrested at G0/G1.Conclusion Bufalin may inhibit the proliferation of QBC939 cells by blocking cell cycle,down-regulating the expression of Cyclin E and up-regulating the expression of P27.