TNP-470对人涎腺腺样囊性癌细胞增殖和凋亡的实验研究

Experimental Study of the Effect of TNP-470 on Human Salivary Adenoid Cystic Carcinoma Cell in Vitro

目的 研究TNP 470对人涎腺腺样囊性癌细胞 (ACC M)的增殖抑制效应及凋亡诱导作用。方法 采用四氮唑盐还原法 (MTT法 )和台盼蓝拒染法 ,检测药物对ACC M细胞的增殖抑制作用 ;荧光染色观察细胞凋亡变化 ;琼脂糖凝胶电泳测定DNA梯状条带 ;流式细胞术检测细胞凋亡峰和细胞周期变化。结果 MTT法和台盼蓝拒染法显示 ,TNP 470对ACC M细胞的半数抑制浓度 (IC5 0 )分别为 40 .6 8μg/ml和 46 .38μg/ml;实验用药组荧光染色观察到细胞呈现凋亡特征 ,DNA电泳显示典型的梯状条带 ;流式细胞术检查有明显的凋亡峰出现 ,细胞周期主要阻滞在G2 /M期 ;实验用药组细胞凋亡率明显增高 ,与对照组有显著性差异 (P <0 .0 1)。结论 TNP 470对ACC M细胞体外直接杀伤作用不明显 ,但可诱导ACC M细胞体外凋亡 ,诱导凋亡可能是其抗肿瘤作用机制之一。

Objective To investigate the effects of TNP 470 on human salivary adenoid cystic carcinoma cell line ACC M in vitro. Methods Cell proliferation was assessed by MTT assays and dye exclusion counting. Morphological changes of apoptosis were observed with fluorescent microscope. DNA ladder, apoptosis rate and cell cycle were examined by DNA agarose gel electronphores and fluorescence flow cytometry (FCM), respectively.Results The 50% inhibitory concentrations (IC50) of TNP 470 on ACC M cells proliferation by MTT assays and dye exclusion counting were 40.68μg/ml and 46.38μg/ml. Apoptosis were observed by fluorescent microscope. DNA electrophoresis for the cells treated with TNP 470 showed brighter DNA ladder; Sub G1 peak and G2/M arrest were also determined by FCM ( P <0.01). Conclusion TNP 470 has the effect of inducing apoptosis in ACC M cells in vitro, which may be one of its antitumor mechanisms. [