汉防己乙素对人肺癌95D细胞凋亡的诱导作用及机制

摘要

Objective To investigate the inducible effect and possible mechanism of fangchinoline on apoptosis of hu-man lung cancer 95D cells. Methods The human lung cancer 95D cells were divided into four groups:experimental groups 1,2,3,and the control group. The cells in the experimental groups 1,2,and 3 were added with 8,16,and 32μmol/ L fangchinoline,respectively. Cells in the control group were added with the same volume of 0. 1% DMSO. The ap-optosis was measured by flow cytometry. The levels of cellular reactive oxygen species (ROS)and Caspase-3 activity were detected by H2 DCF-DA and Caspase-3 activity assay. The BCL-2 and BAX mRNA expression was determined by semi-quantitative RT-PCR. Results The apoptosis rate of the experimental groups 1,2,3 and control group were 6. 71% ± 1. 26%,17. 07% ± 1. 88%,25. 78% ± 0. 44%,and 5. 41% ± 1. 40%,respectively;the relative ROS levels of the ex-perimental groups 1,2,3 and control group were 117. 7% ± 5. 5%,151. 0% ± 4. 6%,212. 0% ± 5. 6%,and 100. 0%± 0. 0%,respectively;the activities of Caspase-3 of the experimental groups 1,2,3 and control group were 119. 3% ± 5. 7%,175. 0% ± 13. 1%,251. 3% ± 17. 0%,and 100. 0% ± 3. 0%,respectively;the levels of BCL-2 mRNA of the ex-perimental groups 1,2,3 and control group were 0. 93 ± 0. 02,0. 75 ± 0. 04,0. 61 ± 0. 04,and 1. 00 ± 0. 03,respective-ly;the levels of BAX mRNA of the experimental groups 1,2,3 and control group were 1. 10 ± 0. 02,1. 63 ± 0. 04,1. 85 ± 0. 04,1. 00 ± 0. 02,respectively. Significant difference was found between the the experimental groups 1,2,3 and the control group (P < 0. 05 or P < 0. 01). Conclusion Fangchinoline may induce the apoptosis of human lung cancer 95D cells by increasing the cellular ROS,activating Caspase-3 and Bax,and inhibiting Bcl-2.%目的 观察汉防己乙素对人肺癌95D细胞凋亡的诱导作用,并探讨其可能的作用机制.方法 取人肺癌95D细胞分为4组,实验1、2、3组分别加入8、16、32μmol/L的汉防己乙素,溶剂对照组加入等体积0.1%的DMSO.采用流式细胞仪测算各组细胞凋亡率,H2DCF-DA法测定细胞内活性氧(ROS)水平,Caspase-3活性测定试剂盒检测Caspase-3活性,半定量RT-PCR法检测BCL-2、BAX mRNA.结果 实验1、2、3组及溶剂对照组细胞凋亡率分别为6.71%±1.26%、17.07%±1.88%、25.78%±0.44%、5.41%±1.40%,ROS水平分别为117.7%±5.5%、151.0%±4.6%、212.0%±5.6%、100.0%±0.0%,Caspase-3活性分别为119.3%±5.7%、175.0%±13.1%、251.3%±17.0%、100.0%±3.0%,BCL-2 mRNA相对表达量分别为0.93±0.02、0.75±0.04、0.61±0.04、1.00±0.03,BAX mRNA相对表达量分别为1.10±0.02、1.63±0.04、1.85±0.04、1.00±0.02,实验1、2、3组与溶剂对照组相比P<0.05或<0.01.结论 汉防己乙素可诱导人肺癌95D细胞凋亡,这可能是通过升高细胞内ROS水平、活化Caspase-3、激活BAX、抑制BCL-2实现的.