雌激素通过NF-κB信号通路影响肝癌细胞的肿瘤学行为研究  

Study on the effect of tumor behavior with estrogen on hepatocellular carcinoma cells through NF-κB signaling pathway

作　　者：李艳萌[1] 贾思雨 徐安健[1] 黄坚[1] LI Yan-meng;JIA Si-yu;XU An-jian(Experimental Research Center,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China;Beijng Clinical Research Institute,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院科研实验中心北京市临床医学研究所,北京100050

出　　处：《临床和实验医学杂志》2019年第2期117-122,共6页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金(编号:81650014)

摘　　要：目的探讨雌激素对HepG2和BEL7402两种肝癌细胞增殖及迁移能力的抑制和诱导凋亡的作用及其相应机制。方法利用MTS试验检测不同浓度的雌激素在不同时间点对HepG2和BEL7402两种肝癌细胞增殖能力的影响。Annexin V/PI流式细胞术检测不同浓度的雌激素诱导HepG2和BEL7402细胞凋亡的情况。Transwell迁移实验检测雌激素对HepG2和BEL7402细胞迁移能力的影响。采用Western blotting法检测不同浓度下雌激素对肝癌细胞中NF-κB信号通路的活性及其下游相关基因c-Myc、Cyclin D1、基质金属蛋白酶(MMP-9)、血管内皮生长因子(VEGF)和Bcl-2表达的影响。结果 MTS试验结果显示,在不同时间点雌激素有效抑制了HepG2和BEL7402细胞的增殖,并随着雌激素浓度的升高其抑制肝癌细胞增殖的效果逐渐增强,与此同时,雌激素也降低了c-Myc、Cyclin D1等增殖相关基因的表达。Annexin V/PI流式细胞术数据显示雌激素诱导HepG2和BEL7402细胞的凋亡并呈明显剂量依赖性,而抗凋亡蛋白Bcl-2的表达出现下降。Transwell迁移实验证明雌激素明显抑制了HepG2和BEL7402细胞的迁移能力并且降低了MMP-9和VEGF的表达。验证了雌激素可有效抑制HepG2和BEL7402肝癌细胞中NF-κB信号通路的活性。结论雌激素通过下调NF-κB信号通路的活性及下游相关分子的表达,而抑制肝癌细胞的增殖与迁移并诱导其凋亡。Objective To explore the role and mechanism of inhibiting proliferation,migration and inducing apoptosis with estrogen on HepG2 and BEL7402 liver cancer cells.Methods MTS assay had been used to detect the effect of estrogen with different concentrations on the capacity of proliferation of HepG2 and BEL7402 liver cancer cells at different time points.Annexin V/PI flow cytometry had been performed to analyze apoptosis of HepG2 and BEL7402 cells induced by estrogen at different concentrations.Transwell migration assay was applied to determine the effect of estrogen on migration capacity of HepG2 and BEL7402 cells.By Western blotting method,the effect of estrogen at different concentrations on the expression of nuclear factor-κB(NF-κB)signaling pathway and its downstream related genes c-Myc,CyclinD1,Matrix metallopeptidase(MMP)-9,vascular endothelial growth factor(VEGF)and Bcl-2 in liver cancer cells were examined.Results Results of MTS assay show that estrogen can effectively inhibit the proliferation of HepG2 and BEL7402 cells in different time points,and following the increase of concentration of estrogen,its inhibition effect on proliferation of liver cancer cells was gradually strengthened.At the same time,estrogen also reduced the expression of c-Myc,CyclinD1 proliferation related genes.The data of Annexin V/PI flow cytometry indicated that estrogen induced apoptosis of HepG2 BEL7402 cells in a significant dosage-dependent manner,and the expression of anti-apoptotic protein Bcl-2 decreased.In addition,Transwell migration assay demonstrated that estrogen can significantly inhibit the migration capacity of HepG2 and BEL7402 cells and decreased the expression of MMP-9 and VEGF.Finally,estrogen can effectively inhibit the activity of NF-κB signaling pathway in HepG2 and BEL7402 liver cancer cells.Conclusion Estrogen inhibits the proliferation and migration of liver cancer cells and induces their apoptosis by down-regulating the activity of NF-κB signaling pathway and the expression of downstream related molecul

关 键 词：肝癌细胞 雌激素 NF-ΚB信号通路 肿瘤学行为 

分 类 号：R735.7[医药卫生—肿瘤]