人肝癌Huh7细胞上皮间充质转化与VEGFa/VEGFR2自分泌途径的相关性及华蟾素的调控作用  

Association Between Epithelial-Mesenchymal Transition and the VEGFa/VEGFR2 Autocrine Pathway in Human Hepatocellular Carcinoma Huh7 Cells and the Regulatory Effect of Cinobufagin

作　　者：叶明君 吴晓红 陈诗雨 李白坤 王萌[2,3] 朱继民[2,4] 李庆林 YE Mingjun;WU Xiaohong;CHEN Shiyu;LI Baikun;WANG Meng;ZHU Jimin;LI Qinglin(School of Pharmacy,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Key Laboratory of Xin an Medicine,Ministry of Education,Anhui Hefei 230012,China;College of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;School of Life Sciences,Anhui University of Chinese Medicine,Anhui Hefei 230012,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]新安医学教育部重点实验室,安徽合肥230012 [3]安徽中医药大学中医学院,安徽合肥230012 [4]安徽中医药大学生命科学学院,安徽合肥230012

出　　处：《安徽中医药大学学报》2024年第5期66-71,共6页Journal of Anhui University of Chinese Medicine

基　　金：国家自然科学基金项目(82305023)。

摘　　要：目的探究华蟾素(cinobufagin,CBG)注射液对人肝癌Huh7细胞血管内皮生长因子a(vascular endothelial growth factor a,VEGFa)/血管内皮生长因子受体2(vascular endothelial growth factor receptor 2,VEGFR2)自分泌信号通路及上皮间充质转化进程的影响。方法通过CCK-8实验、细胞划痕实验、Transwell侵袭实验和成球实验检测人肝癌Huh7细胞增殖、迁移、侵袭和成球能力;免疫荧光双标实验对人肝癌Huh7细胞VEGFa/VEGFR2进行共定位分析,ELISA法检测人肝癌Huh7细胞上清液中VEGFa水平,Western blot法检测人肝癌Huh7细胞中VEGFa/VEGFR2通路及上皮间充质转化相关蛋白表达水平。结果CBG注射液能显著抑制人肝癌Huh7细胞的增殖、迁移、侵袭和成球能力;人肝癌Huh7细胞可能存在VEGFa/VEGFR2自分泌途径;CBG注射液能抑制VEGFa的分泌,降低下游p-VEGFR2、p-PI3K、p-AKT及间充质标志物N-Cadherin、Vimentin、Snail蛋白的表达水平,升高上皮标志物E-Cadherin蛋白的表达水平。结论CBG注射液可能通过VEGFa/VEGFR2自分泌途径调控人肝癌Huh7细胞上皮间充质转化。Objective To investigate the effect of Cinobufagin(CBG)Injection on the VEGFa/VEGFR2 autocrine signaling pathway and epithelial-mesenchymal transition in human hepatocellular carcinoma Huh7 cells.Methods CCK-8 assay,scratch assay,Transwell invasion assay,and sphere-forming assay were used to observe the proliferation,migration,invasion,and sphere formation abilities of Huh7 cells;double-label immunofluorescence was used for the colocalization of VEGFa and VEGFR2 in human hepatocellular carcinoma Huh7 cells;ELISA was used to measure the level of VEGFa in the supernatant of Huh7 cells,and Western blot was used to measure the expression levels of the proteins associated with the VEGFa/VEGFR2 pathway and epithelial-mesenchymal transition in Huh7 cells.Results CBG Injection significantly inhibited the proliferation,migration,invasion,and sphere formation abilities of human hepatocellular carcinoma Huh7 cells.The VEGFa/VEGFR2 autocrine pathway might exist in Huh7 cells,and CBG Injection inhibited the secretion of VEGFa,downregulated the expression levels of downstream p-VEGFR2,p-PI3K,and p-AKT and the mesenchymal markers N-Cadherin,Vimentin,and Snail,and upregulated the protein expression levels of the epithelial marker E-Cadherin.Conclusion CBG Injection may regulate the epithelial-mesenchymal transition of Huh7 cells through the VEGFa/VEGFR2 autocrine pathway.

关 键 词：华蟾素 肝癌 增殖 侵袭 上皮间充质转化 VEGFa/VEGFR2自分泌途径 

分 类 号：R285.5[医药卫生—中药学] R735.7[医药卫生—中医学]