紫草素诱导三阴性乳腺癌细胞MDA-MB-231坏死性凋亡的作用及其机制  

Role and its mechanism of shikonin in induction of necroptosis of triple negative breast cancer MDA-MB-231 cells

作　　者：闫伟平 谢法红[1] 郭瑞杰 聂帅 郭晓辉[1] YAN Weiping;XIE Fahong;GUO Ruijie;NIE Shuai;GUO Xiaohui(Department of Pharmacy,Medical Security Center of Chinese PLA General Hospital,Stationed in Original Outpatient Pharmacy of First Medical Center,Beijing 100853,China)

机构地区：[1]中国人民解放军总医院医疗保障中心药剂科派驻第一医学中心原门诊药局,北京100853

出　　处：《山西医科大学学报》2021年第3期249-254,共6页Journal of Shanxi Medical University

基　　金：军队后勤科研项目重点课题(BWS13C015)。

摘　　要：目的研究紫草素对三阴性乳腺癌细胞MDA-MB-231增殖的影响,并探讨其作用机制。方法配制不同浓度(0,2.5,5,7.5,10,15μmol/L)的紫草素作用于MDA-MB-231细胞,分别培养24,48,72 h,MTT法检测紫草素对细胞存活率的影响;DAPI染色观察细胞核的变化;流式细胞术检测细胞凋亡率。与坏死性凋亡抑制剂Nec-1联用时分为对照组、Nec-1组、紫草素(10μmol/L)组、紫草素与Nec-1联用组;与凋亡抑制剂z-VAD-fmk联用时,分为对照组、z-VAD-fmk组、紫草素(10μmol/L)组、紫草素与z-VAD-fmk联用组;MTT法和流式细胞仪检测细胞OD值和凋亡率的变化,Western blot检测坏死性凋亡关键性蛋白RIP1表达水平的变化。结果紫草素可以明显抑制MDA-MB-231细胞的增殖(P<0.05)。与正常对照细胞相比,紫草素作用后细胞发生皱缩,死亡率增加(P<0.05)。紫草素与Nec-1联用组细胞OD值较紫草素组相比明显增高(P<0.05),细胞凋亡率较紫草素组降低(P<0.05)。而与凋亡抑制剂z-VAD-fmk联用时则几乎没有影响(P>0.05)。RIP1蛋白的表达水平随着紫草素浓度的增加逐渐升高,但与Nec-1联用可以逆转其诱导的RIP1蛋白表达(P<0.05)。结论紫草素可以抑制人三阴性乳腺癌细胞MDA-MB-231的增殖,其作用机制可能与上调RIP1的表达从而诱导坏死性凋亡过程发生有关。Objective To investigate the effect of shikonin on the proliferation of human triple negative breast cancer cell line MDA-MB-231,and explore its mechanism.Methods MDA-MB-231 cells were treated with different concentrations of shikonin(0,2.5,5,7.5,10,15μmol/L)for 24,48,72 h,respectively,MTT assay was used to detect the cell survival rate,DAPI staining was used to observe the changes of cell nucleus,and flow cytometry was used to detect the apoptosis rate.MDA-MB-231 cells were divided into:control group,Nec-1 group,shikonin group(10μmol/L),shikonin+Nec-1 group;or control group,z-VAD-fmk group,shikonin group(10μmol/L),shikonin+z-VAD-fmk group.MTT assay and flow cytometry were used to detect the proliferation and the apoptosis,respectively,and Western blot was used to detect the level of RIP1,a key protein of necroptosis.Results Shikonin significantly inhibited the proliferation of MDA-MB-231 cells(P<0.05).MDA-MB-231 cells became shrinkage after treatment with shikonin,and the apoptosis rate was significantly reduced(P<0.05).Compared with shikonin group,the OD value was significantly increased in shikonin+Nec-1 group(P<0.05),while the apoptosis rate was reduced(P<0.05).But the apoptosis inhibitor z-VAD-fmk had almost no effect on the proliferation and apoptosis(P>0.05).Shikonin increased the expression level of RIP1 protein in a dose-depen-dent manner,and the combination of shikonin and Nec-1 reversed the increase(P<0.05).Conclusion Shikonin can inhibit the proliferation of human triple-negative breast cancer cell MDA-MB-231,and its mechanism may be related to the up-regulation of RIP1 expression to induce the necroptosis.

关 键 词：紫草素 MDA-MB-231 坏死性凋亡 

分 类 号：R739.6[医药卫生—肿瘤]