赖氨大黄酸和紫杉醇对肺癌细胞增殖的抑制作用和凋亡诱导作用

目的:研究赖氨大黄酸（RHL）、紫杉醇单独和两者联合对人肺癌H460细胞增殖和凋亡的影响，阐明其作用机制。方法:选取处于对数生长期肺癌细胞株H460，随机分为对照组(不加药物)、紫杉醇组(1 μmol•L^-1紫杉醇)、RHL组(100 μmol•L^-1 RHL)和紫杉醇联合RHL组，并设空白组。采用MTT法和流式细胞术分别检测各组处理后48 h的细胞增殖率和凋亡率；采用Western blotting 方法检测凋亡相关蛋白和MEK/ERK蛋白的表达水平。结果:细胞培养48 h后，联合用药组细胞增殖率显著低于对照组、紫杉醇组和RHL组 (P<0.05)，联合用药组细胞凋亡率显著高于对照组、紫杉醇组和RHL组(P<0.05)，联合用药组caspase-3和poly ADP-ribose polymerase (PARP)的切割片段蛋白表达强度明显高于对照组、紫杉醇组和RHL组，联合用药组Bcl-2和NF-κB蛋白表达强度明显低于对照组、紫杉醇组和RHL组，同时RHL组紫杉醇上调的MEK和ERK蛋白磷酸化强度降低。结论:紫杉醇和RHL均可抑制肺癌H460细胞增殖，诱导细胞凋亡；RHL通过降低ERK活性、上调caspase-3和PARP的切割片段蛋白表达，增强紫杉醇抑制肺癌细胞增殖和凋亡诱导作用。

Inhibitory effects on proliferation and induction of apoptosis of lung cancer cells of rhein lysinate and taxol

Objective To investigate the effects of rhein lysinate(RHL),taxol and their combination on the proliferation and apoptosis of lung cancer H460 cells,and to clarify their mechanisms.Methods The lung cancer H460 cells in logarithm growth phase were selected and randomly divided into control group(adding no drug),taxol group(1 μmol·L-1taxol),RHL group(100 μmol·L-1RHL) and taxol combined with RHL group;at the same time,blank control group was set up.MTT assay was used to detect the proliferation of H460 cells and flow cytometry method was used to analyze the apoptotic rates in various groups 48 h after treatment.The expression levels of apoptosis-related proteins and MEK/ERK proteins were detected by Western blotting method.Results After culture for 48 h,the cell proliferation rate in taxol combined with RHL group was lower than those in control,taxol,and RHL groups(P<0.05);the apoptotic rate in taxol combined with RHL group was higher than those in control,taxol,and RHL groups(P<0.05);the expressions of caspase-3 and poly ADP-ribose polymerase(PARP) fragment protein in taxol combined with RHL group were higher than those in control,taxol,and RHL groups;whereas the expressions of Bcl-2 and NF-κB proteins were lower than those in control,taxol,and RHL groups.Moreover,the phosphorylation levels of MEK and ERK proteins up-regulated by taxol in RHL group were decreased.Conclusion Both taxol and RHL can inhibit cell proliferation and induce apoptosis of lung cancer H460 cells,and RHL can enhance the taxol-induced cytotoxicity and apoptosis by reducing the activity of ERK and increasing the expression levels of caspase-3 and PARP fragment protein.