紫草素通过上调RIP1、RIP3蛋白诱导下咽癌细胞发生坏死性凋亡  

Shikonin induces necrotic apoptosis in lower pharyngeal cancer Fadu cells by up-regulating RIP1 and RIP3 proteins

作　　者：崔忆旋 赵报 李佳会 柳申成 孙哲[1] 吴俊 丁晓娇 张双双 朱晨露 王楠楠 周兰柱[1] 王文忠[1] CUI Yixuan;ZHAO Bao;LI Jiahui;LIU Shencheng;SUN Zhe;WU Jun;DING Xiaojiao;ZHANG Shuangshuang;ZHU Chenlu;WANG Nannan;ZHOU Lanzhu;WANG Wenzhong(Department of Otorhinolaryngology Head and Neck Surgery,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院耳鼻咽喉科

出　　处：《山西医科大学学报》2019年第11期1527-1530,共4页Journal of Shanxi Medical University

基　　金：安徽省高校自然科学研究重点项目(KJ2019A0293,KJ2019A0400);蚌埠医学院科技发展基金项目(BYKF1763);蚌埠医学院自然科学基金重点项目(BYKY1855ZD)

摘　　要：目的探究紫草素对下咽癌细胞(Fadu)的增殖和死亡的影响。方法不同浓度的紫草素(0,2,4,6,8,10μmol/L)分别处理Fadu细胞24,48,72 h,检测细胞的存活率,其中0μmol/L组为空白对照组。MTT法检测紫草素对Fadu细胞增殖的影响;AnnexinⅤ-FITC/PI双染法检测不同浓度紫草素(0,2,4,6μmol/L)作用于Fadu细胞24 h的凋亡率;Western blot法检测紫草素作用Fadu细胞后Bcl-2、Bax、RIP1、RIP3蛋白相对表达水平。结果 MTT结果显示,紫草素对Fadu细胞的增殖抑制作用呈时间依赖性及剂量依赖性(P <0. 05),24,48,72 h的IC50分别为6. 012,4. 120,3. 939μmol/L;AnnexinⅤ-FITC/PI双染法可知紫草素可随浓度依赖性诱导Fadu细胞凋亡;Western blot结果显示,紫草素(2,4,6μmol/L)可以下调Bcl-2和上调Bax,诱导Fadu细胞凋亡,上调RIP1、RIP3蛋白诱导下咽癌细胞坏死性凋亡。结论紫草素可诱导Fadu细胞的凋亡和坏死性凋亡,凋亡机制可能为下调凋亡抑制蛋白Bcl-2和上调凋亡诱导蛋白Bax的表达,坏死性凋亡的机制可能为上调RIP1、RIP3蛋白的表达。Objective To investigate the effects of shikonin on the proliferation and death of hypopharyngeal carcinoma cells( Fadu).Methods Fadu cells were treated with different concentrations of shikonin( 0,2,4,6,8,10 μmol/L) for 24,48,72 h,respectively. The viability of the cells was detected,and the 0 μmol/L group was chosen as blank control group. The effect of shikonin on the proliferation of Fadu cells was detected by MTT assay. The apoptosis rate of Fadu cells was detected by Annexin Ⅴ-FITC/PI double staining after treated with different concentrations of shikonin( 0,2,4,6 μmol/L) for 24 h. Western blot was used to detect the relative expression levels of Bcl-2,Bax,RIP1 and RIP3 proteins in Fadu cells treated with shikonin. Results The results of MTT showed that the shikonin inhibited the proliferation of Fadu cells in a time-dependent and dose-dependent manner( P < 0. 05). The IC50 values at 24,48 h and 72 h were 6. 012,4. 120 and 3. 939 μmol/L,respectively. Annexin Ⅴ-FITC/PI double staining showed that shikonin induced apoptosis of Fadu cells in a concentration-dependent manner. Western blot analysis showed that shikonin( 2,4,6μmol/L) down-regulated Bcl-2 and up-regulated Bax,induced the apoptosis of Fadu cells and up-regulated RIP1 and RIP3 proteins to induce necrotic apoptosis of hypopharyngeal carcinoma cells. Conclusion Shikonin may induce apoptosis and necrotic apoptosis of Fadu cells. The mechanism of apoptosis may be related to down-regulation of apoptosis-inhibiting protein Bcl-2 and up-regulation of apoptosisinducing protein Bax. The mechanism of necrotic apoptosis may be related to up-regulation of the RIP1,RIP3 protein expression.

关 键 词：紫草素 下咽癌 Fadu细胞 坏死性凋亡 

分 类 号：R739.63[医药卫生—肿瘤]