大黄素下调ERCC1和Rad51对非小细胞肺癌增殖的影响及分子机制  

Emodin down-regulates ERCC1 and Rad51 and inhibits proliferation in non-small cell lung cancer cells

作　　者：何林[1] 毕娟娟[1] 郭倩[1] 余音[1] 叶秀峰[1] 李伟 

机构地区：[1]重庆医科大学基础医学院病理学教研室,重庆市神经生物学重点实验室,重庆400016

出　　处：《第三军医大学学报》2011年第22期2370-2375,共6页Journal of Third Military Medical University

基　　金：重庆市科技攻关计划项目(CSTC2008AB5118)~~

摘　　要：目的研究大黄素对非小细胞肺癌的细胞毒性,并观察其对非小细胞肺癌ERCC1和Rad51表达的影响。方法培养肺腺癌细胞株A549和肺鳞癌细胞株SK-MES-1,据MTT法测定结果将细胞分为5组:①对照组;②大黄素处理组;③溶剂处理组;④U0126处理组;⑤大黄素联合U0126处理组。细胞以不同浓度的大黄素处理48 h后分为4组。用RT-PCR和Western blot法测定目的基因RECC1和Rad51的表达情况,透射电镜观察两种细胞在大黄素作用前后的变化。结果 MTT法测定出大黄素作用48 h时A549和SK-MES-1细胞株IC50值分别为(70.18±1.94)μmol/L和(41.95±1.27)μmol/L。大黄素100μmol/L处理A549和SK-MES-1 48 h后抑制率为(60.42±1.37)%和(75.48±0.48)%,明显高于其他组(P<0.05)。RT-PCR和Western blot检测结果显示:大黄素作用A549和SK-MES-1浓度越大,ERCC1和Rad51表达水平越低,且差异具有统计学意义(P<0.05)。大黄素与ERK信号通路阻滞剂U0126联合使用时,ERCC1和Rad51表达明显低于大黄素和U0126单独使用(P<0.05)。大黄素对A549和SK-MES-1细胞均具有生长抑制作用,其细胞毒性呈浓度依赖性。电镜观察可见大黄素处理的A549和SK-MES-1细胞有空泡变性。结论大黄素可降低ERCC1和Rad51的表达,从而引起非小细胞肺癌细胞生长抑制。Objective To study emodin-mediated cytotoxicity and determine the effect of emodin on Rad51 and ERCC1 expression in non-small cell lung cancer（NSCLC）.Methods A549 cells and SK-MES-1 cells were cultured and their viability was evaluated by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay.Then the cells were treated by emodin,menstruum,U0126（ERK signal passway inhibitor）,or emodin＋U0126 respectively.After the cells were treated with emodin at different concentrations for 48 h,the mRNA and protein levels of ERCC1 and Rad51 were determined by RT-PCR and Western blot analysis.The ultrastructure of A549 and SK-MES-1 cells were observed by transmission electron microscopy before and after emodin treatment.Results After emodin treatment for 48 h,MTT assay showed that the IC50 values of A549 cells and SK-MES-1 cells were 70.18±1.94 and 41.95±1.27 μmol/L,respectively.Cell proliferation inhibition ratio in A549 and SK-MES-1 cells were（60.42±1.37）% and（75.48±0.48）% after 100 μmol/L emodin treatment for 48 h,respectively,and were significantly higher than those in cells after other treatment（P0.05）.RT-PCR and Western blot analysis showed that emodin inhibited the mRNA and protein levels of ERCC1 and Rad51 in A549 and SK-MES-1 cells in a dose dependent manner（P0.05）.The combination of emodin and U0126 had a stronger effect on inhibiting the expressions of Rad51 and ERCC1 when compared with single drug at the same dose（P0.05）.Emodin mediated cytotoxicity in NSCLC was in a dose dependent manner.Transmission electron microscopy displayed vacuolar degenerations were observed in A549 and SK-MES-1 cells after emodin treatment.Conclusion Emodin inhibits cell proliferation in NSCLC by downregulating ERCC1 and Rad51.

关 键 词：大黄素 ERCC1 RAD51 U0126 空泡变性 

分 类 号：R285.5[医药卫生—中药学] R730.23[医药卫生—中医学]