多西他赛对三阴性乳腺癌细胞株增殖影响及其机制的探讨  

Effect of Docetaxel on the proliferation of triple negative breast cancer cell line and its mechanism

作　　者：查全斌[1] 唐金海[2] 季明华[3] 吴建中[4] 乔恩奇[5] 何跃君[6] 

机构地区：[1]徐州医学院肿瘤学教研室,江苏徐州221004 [2]江苏省肿瘤医院乳腺外科,江苏南京210009 [3]江苏省肿瘤医院放疗科,江苏南京210009 [4]江苏省肿瘤医院科研科,江苏南京210009 [5]南京医科大学肿瘤学系,江苏南京210009 [6]徐州医学院外科学教研室,江苏徐州221004

出　　处：《中华肿瘤防治杂志》2012年第23期1761-1766,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30840093);江苏省财政厅和科技厅资助项目(BM2011118)

摘　　要：目的:研究多西他赛(Docetaxel)对三阴性乳腺癌细胞株MDA-MB-231增殖抑制作用,探讨作用过程中与MAPK信号转导通路的关系及相关机制。方法:采用CCK-8试剂盒检测多西他赛对MDA-MB-231细胞增殖的抑制作用;倒置显微镜观察肿瘤细胞的形态学变化;流式细胞术(FCM)分析多西他赛作用后细胞周期分布及凋亡情况;蛋白质印迹法分别检测多西他赛处理后MAPK通路蛋白和凋亡相关蛋白Bcl-2、Bax的表达。结果:多西他赛可抑制MDA-MB-231细胞增殖,呈浓度、时间依赖性。显微镜观察肿瘤细胞形态发生明显改变,悬浮细胞增多。流式细胞术分析可见,在多西他赛1.25和2.50μg/mL浓度下,停滞于G2/M期细胞百分率分别为(16.52±1.30)%和(29.15±0.54)%,明显高于对照组的(6.13±0.59)%,P<0.01;同时细胞凋亡率分别为(2.61±0.09)%和(3.15±0.07)%,明显高于对照组的(0.42±0.02)%,P<0.05。蛋白质印迹法结果显示,多西他赛处理后,细胞p-ERK1/2蛋白的表达水平下降,p-JNK/SAPK蛋白的表达水平上升,而总ERK1/2、JNK/SAPK、p38及p-p38蛋白的表达水平无明显变化。凋亡相关蛋白Bcl-2的表达降低,Bax的表达升高。结论:多西他赛通过影响ERK1/2和JNK/SAPK信号转导通路,调节凋亡相关蛋白的表达而抑制三阴性乳腺癌细胞株MDA-MB-231的增殖。OBJECTIVE： To study the inhibitory effects of docetaxel on the proliferation of triple negative breast cancer line MDA-MB-231 in vitro, and investigate the related mechanism with MAPK singal transduction pathway. METHODS： The inhibitory effects of docetaxel on the proliferation of MDA-MB-231 ceils were caculated by cell countiny kit-8 assay. The morphological changes of cells were observed with inverted microscope. Flow cytometry was used to de termine cell cycle distribution and apoptosis,and Western blot was used to detect the protein expression level changes of MAPK pathway members,Bcl-2 and Bax when cells were given Docetaxel treatment. RESULTS： Docetaxel inhibited the proliferation of MDA-MB-231 cells in a time-and concerntration-dependent manner. The morphology of tumor cells has changed significantly and the number of suspension cells were increased under microscope. The result of flow cytometry indicated that at 1.25,2.50/~g/mL docetaxel,the percentage of Gz/M cell was （16.52±1.3）% and （29.15±0. 54）% re spectively,much higher than that of the control（6.13 ± 0.59）% （P〈 0.01）% meanwhile, the apoptosis rate of daeetaxel group were （2.61±0.09）% and （3.15± 0.07）% respectively, much higher than that contrast group （0, 42±0.02） （P〈0.05）. Docetaxel down-regulated the expression level of p-ERK1/2 and Bcl-2,while up-regulated that of p-JNIK/ SAPK and Bax in MDA-MB-231 cell. However, there was no marked change in the expression of total ERK1/2 JNK/SAPK, p38 and p-p38. CONCLUSION： Docetaxel inhibits cell proliferation in vitro by affecting the ERK1/2,JNK/SAPK singal transduetion pathway and regulating the expression of apoptosis related proteins in triple negative breast cancer cell line MDA-MB 231.

关 键 词：乳腺肿瘤 多西他赛 ERK1/2 JNK/SAPK MAPK信号传导 三阴性 印迹法 蛋白质 

分 类 号：R737.9[医药卫生—肿瘤]