左旋棉酚诱导daudi细胞发生自噬的可能机制及 - 第三军医大学学报.doc

左旋棉酚通过ERK通路诱导Daudi细胞发生自噬及意义 倪振洪，王滨，丁雯，程攀科，连继勤*，何凤田* （400038重庆，第三军医大学基础医学部生物化学与分子生物学教研室） 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；AO染色后细胞内可观察到的Western blot显示左旋棉酚上调自噬蛋白LC3Ⅱ的表达；自噬抑制剂左旋棉酚。结论 关键词：左旋棉酚；Burkitt淋巴瘤；细胞增殖；自噬；ERK The mechanism and significance of (-)-gossypol-induced Autophagy in Daudi cells Ni Zhenhong, Wang Bin, Ding Wen, Cheng Panke, Lian Jiqin*, He Fengtian* Department of Biochemistry and Molecular Biology, Third Millitary Medical University, Chongqing, 400038 China Abstract：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 proliferation in Daudi cells in vitro. Trypan blue exclusion assay was used to detect cell viability during different treatments. Western blot was used to determine the expression of LC3 II and ERK. Acridine Orange staining was used to detect the formation of acidic vesicular organelles. 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 revealed that (-)-gossypol treatment markedly upregulated LC3 II and induced phosphorylation of ERK. Inhibition of ERK activity may block upregulation of LC3 II mediated by (-)-gossypol. Inhibition of autophagy by U0126,CQ and 3-MA can enhance cell death mediated by (-)-gossypol. Conclusion (-)-gossypol can effectively inhibit cell proliferation and induce autophagy in Burkitt lymphoma Daudi cells via ERK pathway. . Key words：(-)-gossypol;Burkitt lymphoma;cell proliferation;autophagy Grand support: Chongqing Natural Science Foundation (CSTC, 2011BB5030) Corresponding author: Lian Jiqin, E-mail: lianjiqin@；He Fengtian，E-mail: hefengtian06@ 基金项目：重庆市自然科