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槐耳通过诱导自噬抑制肝癌细胞增殖与迁移 预览 被引量:1

Huaier inhibits proliferation and metastasis of liver cancer cells by inducing autophagy
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摘要 目的探讨槐耳对人肝细胞癌(肝癌)细胞增殖与迁移的影响,并探讨其相关分子机制。方法将肝癌细胞株SK-HEP-1和HEP3B分别设为对照组、槐耳处理组和自噬抑制剂组。CCK-8方法检测细胞活性,克隆形成实验检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell实验检测细胞迁移,GFP-LC3荧光融合蛋白检测自噬小体的形成,Westernblot检测上皮-间充质转化(EMT)、自噬和Akt/mTOR通路相关蛋白。两组数据比较采用t检验,多组比较采用单因素方差分析和LSD-t检验。结果肝癌细胞SK-HEP-1与HEP3B活性随着槐耳浓度升高与时间的增加而降低。6g/L槐耳处理组肝癌细胞SK-HEP-1和HEP3B的克隆数分别为(77±9)、(84±9)个,明显少于对照组的(231±15)、(257±35)个(t=-8.90,-6.33;P<0.05)。槐耳可明显促进细胞凋亡,具有浓度依赖性(F=120.88,199.59;P<0.05)。8g/L槐耳处理组肝癌细胞SK-HEP-1和HEP3B的穿膜细胞数分别为(68±13)、(54±13)个,明显少于对照组的(389±28)、(411±15)个(t=-17.98,-31.47;P<0.05)。Westernblot检测显示,槐耳处理组N-cadhenin和Vimentin蛋白表达减弱;LC3Ⅱ表达增强,LC3Ⅰ表达减弱;p-Akt、p-mTOR、p62蛋白表达减弱,同时诱导LC3Ⅱ表达增强。自噬抑制剂3-MA可减少槐耳处理组的自噬。结论槐耳通过促进肝癌细胞凋亡及抑制细胞增殖和转移而发挥抗肿瘤效应,其机制可能与抑制Akt/mTOR通路诱导自噬和阻止EMT有关。 Objective To evaluate the effect of Huaier on the proliferation and metastasis of human hepatocellular carcinoma (HCC) cells, and to explore the molecular mechanism. Methods HCC cell lines SK-HEP-1 and HEP3B were divided into the control, Huaier treatment and autophagy inhibitor groups. The cell viability was detected by CCK-8 assay. The cell proliferation was determined by clone formation assay. The cell apoptosis was detected by flow cytometry. The cell metastasis was detected by Transwell assay. The formation of autophagosome was observed by fluorescent GFP-LC3 fusion protein.The related proteins of epithelial-mesenchymal transition (EMT), autophagy and Akt/mTOR signaling pathway were detected by Western blot. Comparison between 2 groups was conducted by t test. Multi-group comparison was carried out by One-way ANOVA and LSD-t test. Results The activity of SK-HEP-1 and HEP3B decreased with the increase of Huaier concentration and the prolongation of treatment time. In 6 g/L Huaier treatment group, the number of clone formation of SK-HEP-1 and HEP3B cells was 77±9 and 84±9 respectively, significantly less than 231±15 and 257±35 in control group (t=-8.90,-6.33;P<0.05). Huaier significantly promoted the cell apoptosis in a concentration-dependent manner (F=120.88, 199.59;P<0.05). In 8 g/L Huaier treatment group, the number of trans-membrane SK-HEP-1 and HEP3B cells was 68±13 and 54±13, significantly less than 389±28 and 411±15 in control group (t=-17.98,-31.47;P<0.05). Western blot demonstrated that the expression of N-cadherin and Vimentin proteins was down-regulated, LC3Ⅱ expression was up-regulated and LC3Ⅰ expression was down-regulated in the Huaier treatment group. The expression levels of p-Akt, p-mTOR and p62 proteins were down-regulated and the expression of LC3Ⅱ was up-regulated. Autophagy inhibitor 3-MA could reduce the autophagy in Huaier treatment group. Conclusions Huaier exerts anti-tumor effect by promoting the apoptosis and inhibiting the proliferation and metastasis of HCC cells. Th
作者 黎文信 王喜城 向美焕 谭文亮 徐云修修 叶义标 陈捷 陈涛 Li Wenxin;Wang Xicheng;Xiang Meihuan;Tan Wenliang;Xu Yunxiuxiu;Ye Yibiao;Chen Jie;Chen Tao(Department of Hepatobiliary Surgery,Sun Yat-sen Memorial Hospital,Sun Yat-senUniversity,Guangzhou 510120,China;Guangdong Provincial Key Laboratory of Malignant TumorEpigenetics and Gene Regulation,Sun Yat-sen Memorial Hospital,Sun Yat-senUniversity,Guangzhou 510120,China;Nursing Department,Sun Yat-sen Memorial Hospital,Sun Yat-senUniversity,Guangzhou 510120,China)
出处 《中华肝脏外科手术学电子杂志》 CAS 2019年第2期169-174,共6页 Chinese Journal of Hepatic Surgery(Electronic Edition)
基金 国家自然基金青年项目(81800560) 广州市科技产学研项目(201604030054).
关键词 肝细胞 自噬 信号通路 细胞增殖 细胞运动 Carcinoma, hepatocellular Autophagy Signaling pathway Cell proliferation Cell movement
作者简介 通信作者: 陈涛,Email:ct0498@126.com
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