miR-200c增强肝星状细胞促肝细胞癌生长的能力  

Hepatic stellate cell accelerates the development of hepatocellular carcinoma through miR-200c

作　　者：蔡秀琴[1] 吴健[1] 马腾飞[1] 李巧[1] 华赟鹏[1] CAI Xiu-qin;WU Jian;MA Teng-fei;LI Qiao;HUA Yun-peng(Department of Hepatic Surgery, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, China)

机构地区：[1]中山大学附属第一医院肝外科,广东广州510080

出　　处：《热带医学杂志》2018年第6期712-716,F0003,共6页Journal of Tropical Medicine

基　　金：广东省科技计划项目(2014A020212626)

摘　　要：目的探讨mi R-200c活化的肝星状细胞(HSC)对肝细胞癌(HCC)的促进作用。方法用慢病毒转染的方法构建过表达mi R-200c的HSC稳定株(LX2-200c)以及空质粒稳定株(LX2-nc);用MTT实验检测细胞增殖能力的变化,用transwell小室检测迁移和侵袭能力的影响;用裸鼠皮下种植瘤观察肝星状细胞对肝癌细胞株(Hep G2和Hep3B)成瘤的影响;用Western blot方法检测Hep G2和Hep3B内上皮间质转化(EMT)的标记物Zeb-1、N-cadherin和Vemintin的表达;用免疫组化方法检测皮下种植瘤组织内肝星状细胞活化标志物(α-SMA)、肿瘤组织增殖状态标志物(Ki67)的表达水平。结果过表达mi R-200c的HSC细胞株(LX2-200c)的条件培养基与对照组(LX2-nc)的条件培养基相比,可显著促进HCC体外增殖(P<0.05)、迁移及侵袭。LX2-200c可促进肝癌细胞皮下成瘤的重量和体积(P<0.05),并促进种植瘤组织中Ki67及α-SMA表达显著增加。LX2-200c条件培养基作用后的肝癌细胞株上Zeb-1、N-cadherin和Vemintin的表达增加。结论 mi R-200c活化的肝星状细胞可显著促进肝癌细胞生长、侵袭。Objective This study aims to investigate the role of mi R-200 c in hepatic stellate cell（HSC）accelerating the growth of hepatocellular carcinoma（HCC）.Methods HSC cell lines stably express mi R-200 c（LX2-200 c）or blank plasmid（LX2-nc）was constructed by lentiviral transfection.Cell proliferative ability was detected by MTT assay.Migration and invasion of cells were detected by transwell chamber assay.The subcutaneous xenograft tumors in nude mice were used to observe the influence of HSCs on the development of HCC（Hep G2 and Hep3 B）.The expression of Zeb-1,N-cadherin and Vemintin,the markers of epithelial-mesenchymal transition（EMT）in Hep G2 and Hep3 B,were measured by Western blot.The marker of activated HSC（α-SMA） and a tumor cellular marker for proliferation（Ki67） were detected by immunostaining.Results The conditioned medium of HSC cell line stably expressing mi R-200 c（LX2-200 c）significantly promoted the proliferation（P〈0.05）,migration and invasion of HCC.Co-culture of LX2-200 c and HCC cell lines can accelerate weight and volume of xenografts in nude mice（P〈0.05）.In addition,we found the high levels of Zeb-1,Ncadherin and Vemintin in HCC cell lines treated with the conditional medium of LX2-200 c.The levels of α-SMA and Ki67 in the xenografts from the co-culture of LX2-200 c and HCC cell lines were significantly higher than LX2-nc control.Conclusion HSC could promote the growth and invasion of HCC through mi R-200 c.

关 键 词：MIR-200C 肝星形细胞 肝细胞癌 

分 类 号：R735.7[医药卫生—肿瘤]