左旋棉酚通过ERK通路诱导Daudi细胞发生自噬及意义  

(-)-gossypol induces autophagy in Daudi cells through ERK signal pathway

作　　者：倪振洪[1] 王滨[1] 丁雯[1] 程攀科[1] 连继勤[1] 何凤田[1] 

机构地区：[1]第三军医大学基础医学部生物化学与分子生物学教研室,重庆400038

出　　处：《第三军医大学学报》2012年第23期2384-2387,共4页Journal of Third Military Medical University

基　　金：重庆市自然科学基金(CSTC2011BB5030)~~

摘　　要：目的探讨左旋棉酚诱导淋巴瘤Daudi细胞发生自噬可能机制及对细胞存活率的影响。方法采用CCK-8法检测左旋棉酚在体外对Daudi细胞增殖抑制作用的影响;采用台盼蓝排斥实验检测不同处理对细胞存活率的影响;Western blot检测细胞中自噬相关蛋白LC3、ERK和磷酸化ERK的表达情况;AO染色观察经左旋棉酚处理后Daudi细胞酸性小体的变化情况。结果左旋棉酚能剂量依赖性地抑制Daudi细胞的增殖和促进细胞死亡;AO染色后经左旋棉酚处理的细胞内可观察到大量的酸性小体形成;Western blot显示左旋棉酚能显著上调自噬相关蛋白LC3Ⅱ的表达及增强磷酸化ERK的水平,抑制ERK的磷酸化能下调LC3Ⅱ的表达;ERK抑制剂U0126及自噬抑制剂CQ和3-MA均能显著增强左旋棉酚的杀细胞效力。结论左旋棉酚可能通过ERK通路诱导Daudi细胞发生自噬,抑制ERK介导的自噬能够显著增强左旋棉酚的抗瘤效应。Objective To investigate the mechanism of （-）-gossypol-induced autophagy and its effect on cell viability in Burkitt lymphoma Daudi cells. Methods CCK-8 detection was used to assess the inhibitory effects of （-）-gossypol on the proliferation in Daudi cells. Trypan blue exclusion assay was used to detect cell viability during different treatments. Western blotting was used to determine the expression of LC3 Ⅱ and ERK. Acridine orange staining was used to detect the formation of acidic vesicular organelles （AVO）. Results （-）-gossypol inhibited cell proliferation and induced cell death in a dose-dependent manner. Increased AVOs were noted after treatment of cells with （-）-gossypol. Western blot analysis revealed that （-）-gossypol treat- ment markedly upregulated LC3Ⅱ and induced phosphorylation of ERK. Inhibition of ERK activity blocked the upregulation of LC3 Ⅱ mediated by （-） -gossypol. Inhibition of autophagy by U0126 （ ERK inhibitor）, CQ and 3-MA （autophagy inhibitors） enhanced cell death mediated by （-）-gossypol. Conclusion （-）-gossypol effectively inhibits cell proliferation and induces autophagy in Burkitt lymphoma Daudi cells via ERK pathway.

关 键 词：左旋棉酚 BURKITT淋巴瘤 细胞增殖 自噬 ERK 

分 类 号：R733.4[医药卫生—肿瘤] R965[医药卫生—临床医学]