热化联合对人小细胞肺癌H446细胞凋亡的影响

Effect of thermo-chemo therapy on human small cell lung cancer H446 cell apoptosis

目的研究热化联合对肺部肿瘤细胞生长的影响及其可能的机制。方法参考临床常用剂量,采用43℃加热联合120μg/L紫杉醇(热化联合组)、43℃加热联合120μg/L紫杉醇并加入30μmol/L活性氧(ROS)特异抑制剂NAC(NAC组)、单纯使用120μg/L紫杉醇(单纯化疗组)处理H446细胞,以未处理的H446细胞作对照,应用流式细胞术(FCM)检测细胞凋亡率,荧光法检测细胞内活性氧(ROS),蛋白免疫印记法(Western blotting)检测Caspase-3的表达,SPSS 13.0对数据进行统计分析。结果热化联合组细胞凋亡率高于其余各组(P<0.05);ROS在热化联合组增高(P<0.05),NAC可抑制其表达;Caspase-3表达在热化联合组中表达明显增高(P<0.05),但可被NAC抑制(P<0.05)。结论热化联合应用可以明显诱导H446细胞发生凋亡,可能是通过诱导ROS的产生,通过caspase途径实现的。

Objective To study the synergistic effect of thermo-chemo therapy on lung cancer cells and its possible mechanisms.Methods Refer the normal clinic dose,and use the untreated H446 cells as the control group.H446 cells were treated by different thermo-chemo therapy strategies： 43℃ ＋ Paclitaxel（120μg /L）（thermo-chemotherapy group）,43℃ ＋ Paclitaxel（120μg/L） ＋ NAC（30μmol/L,specific reactive oxygen species（ROS） inhibitor）（NAC group）,and Paclitaxel（120μg /L） alone（only chemotherapy group）.Cell apoptosis was analyzed by FCM and fluorescence was used to measure ROS inside the cells.The expressions of Caspase-3 was determined by Westerm Blotting.Use SPSS 13.0 to perform the statistical analysis on the data.Results The rate of cell apoptosis in the thermo-chemo therapy group was significantly higher than those in the other groups（P〈0.05）.The ROS in the thermo-chemo therapy group increased（P〈0.05）,but NAC inhibited its expression.The expression of Caspase-3 in the thermo-chemo therapy group increased significantly（P〈0.05）,but can be inhibited by NAC（P〈0.05）.Conclusion Thermo-chemo therapy can significantly seduce the apotosis of H446 cells,probably through seducing ROS forming,realised by the caspase channel.