热化疗联合作用抑制人小细胞肺癌细胞增殖的机制

Mechanism of Inhibition of Thermo-Chemotherapy on Cell Proliferation of Human Small Cell Lung Cancer

目的研究热化疗联合作用对肺部肿瘤细胞生长的抑制及其可能的机制。方法参考临床常用剂量,采用43℃加热联合120μg/L紫杉醇、43℃加热联合120μg/L紫杉醇及20μmol/LJNK特异抑制剂SP600125以及单纯使用120μg/L紫杉醇处理H446细胞,以未处理的H446细胞作对照。应用噻唑蓝比色法检测各处理方式下细胞增殖率的变化,通过蛋白免疫印迹法检测JNK磷酸化水平和Caspase-3表达的变化,流式细胞术检测细胞凋亡率,运用SPSS13.0对数据进行统计分析。结果热化联合组细胞增殖率最低(P<0.05);JNK磷酸化水平在热化联合组中表达明显增高(P<0.05),SP600125可抑制其磷酸化(P<0.05);热化联合组的Caspase-3增高(P<0.05),SP600125使其表达减少(P<0.05);热化联合组细胞凋亡率增加(P<0.05),SP600125组细胞凋亡率减少(P<0.05)。结论热化疗联合应用可以明显增加紫杉醇对H446细胞生长的抑制作用,且这种抑制作用可能是通过激活JNK信号转导通路,继而通过Caspase-3途径激活细胞凋亡来实现的。

Objective To study the inhibition of thermo-chemotherapy on lung cancer and its possible mechanism.Methods H446 cells were subjected to different thermo-chemotherapy strategies：43 ℃＋Paclitaxel（120 μg/L）,43 ℃＋Paclitaxel（120 μg/L）＋SP600125（20 μmol/L,JNK inhibitor）,and Paclitaxel（120 μg/L）.The untreated cells were served as control.MTT assay was used to measure cell proliferation and Western blot was used to examine the phosphorylation of JNK and the expression of Caspase-3.Results The proliferation rate of cells in the thermo-chemotherapy group was significantly lower than those in the other groups（P0.05）.The phosphorylation of JNK significantly increased in the thermo-chemotherapy group（P0.05）;SP600125 inhibited the phosphorylation of JNK and the proliferation of cells in the SP600125 group elevated（P0.05）.The expression of Caspase-3 in the thermo-chemotherapy group increased,which could be inhibited by SP600125（P0.05）.The rate of apoptosis increased in the thermo-chemotherapy group（P0.05） and decreased in SP600125 group（P0.05）.Conclusion Thermotherapy can promote the inhibition effect of Paclitaxel chemotherapy against the growth of lung cancer cell line H446,probably through activating JNK pathway and Caspase-3 to induce apoptosis.