异泽兰黄素对肾癌786—O细胞增殖的抑制作用及分子机制  

Antitumor activity of Eupatilin in renal cell carcinoma 786-O cell line and its mechanism

作　　者：钟伟枫[1,2] 刘思平[1] 陈南辉[1] 万沛[1] 林毅锋[1] 江惠明[1] 钟凯华 赵平森[1] 周芳坚[2] Zhong Weifeng;Liu Siping;Chen Nanhui;Wan Pei;Lin Yifeng;Jian Huiming;Zhong Kaihua;Zhao Pinseng;Zhou Fangjian(Department of Urology,Meizhou People＇s Hospital,Meizhou 514021,China;Department of Urology;Sum Yat-sen Cancer Center,Guangzhou 510060,Chin)

机构地区：[1]梅州市人民医院泌尿外科,514021 [2]中山大学肿瘤防治中心泌尿外科,广州510060

出　　处：《国际医药卫生导报》2018年第16期2403-2408,2416,共7页International Medicine and Health Guidance News

基　　金：广东省自然科学基金-粤东西北创新人才联合培养项目（2017A030307037）;中国博士后科学基金项目（2016M602595）

摘　　要：目的 探讨异泽兰黄素对肾癌786-O细胞增殖的影响及分子机制.方法 应用MTT法检测不同浓度（2.5、5.0、10.0、20.0、40.0 mmol/L）异泽兰黄素对肾癌786-O细胞活力的影响,流式细胞仪检测异泽兰黄素对肾癌786-O细胞凋亡的影响,Transwell检测异泽兰黄素对肾癌786-O细胞侵袭的影响,荧光定量PCR检测异泽兰黄素对肾癌786-O细胞miR-21表达的影响.转染病毒构建miR-21稳定表达细胞株,MTT、流式细胞仪和Transwell分别检测miR-21和异泽兰黄素对肾癌786-O细胞增殖、凋亡和侵袭的影响,Western blot法检测miR-21和异泽兰黄素对细胞内p-AKT、AKT、Bax和Bcl2蛋白表达的影响.结果 2.5、5.0、10.0、20.0、40.0 mmol/L异泽兰黄素作用肾癌786-O细胞,细胞活力显著降低,且呈现浓度依赖性.与正常对照组相比,异泽兰黄素作用细胞24 h能显著促进细胞凋亡（P＜0.01）,抑制细胞侵袭（P＜0.01）,抑制miR-21的表达（P＜0.01）,且作用均具有浓度依赖性;Western blot结果显示,异泽兰黄素抑制AKT磷酸化水平和Bcl2蛋白表达,促进Bax蛋白表达,与正常对照组相比意义具有显著统计学差异.过表达miR-21能显著抑制异泽兰黄素诱导的细胞凋亡和侵袭,并抑制异泽兰黄素调控的p-AKT、Bax和Bcl2的表达.结论 异泽兰黄素可抑制肾癌786-O细胞miR-21的表达,通过激活AKT通路诱导细胞凋亡.Objective To investigate the molecular mechanism of Eupatilin in inhibiting the proliferation of renal cell carcinoma （RCC）.Methods The influence of Eupatilin on the viability of RCC was evaluated by MTT assay.The apoptosis induced by Eupatilin was measured by flow cytometry analysis.The invasion of cells induced by Eupatilin was measured by Transwell.The miR-21 level was measured by Q-PCR.After overexpression of miR-21,the effects of miR-21 and Eupatilin on the proliferation,apoptosis,and invasion of human renal cell carcinoma 786-O cells were detected by MTT,flow cytometry,and Transwell,respectively.The protein levels of AKT,p-AKT,Bax,and Bcl2 were examined by western blot analysis.Results 2.5,5.0,10.0,20.0 and 40.0 mmol/L Eupatilin could significantly inhibit the viability of RCC in a dose-dependent manner.Eupatilin could significantly promote the apoptosis of 786-O,and inhibit cell invasion and expression of miR-21 in a dosedependent manner.Western blot results showed that Eupatilin inhibited the expressions of p-AKT and Bcl2 and promoted the expression of Bax,which was significantly different from that of the control group.The overexpression of miR-21 significantly inhibited Eupatilin-induced apoptosis and Eupatilin-regulated expressions of p-AKT,Bax,and Bcl2.Conclusion Eupatilin inhibits miR-21 expression in renal carcinoma 786-O cells and induces apoptosis by activating AKT pathway.

关 键 词：异泽兰黄素 肾癌786-O细胞 细胞凋亡 AKT通路 

分 类 号：R285[医药卫生—中药学]