活化的肝星状细胞通过CCL22/CCR4轴促进肝癌细胞侵袭能力  

Activated hepatic stellate cells induce hepatocellular carcinoma invasion through CCL22/CCR4 axis

作　　者：王霆[1] 徐凌[1] 魏珏[1] 王玉刚[1] 惠萍萍[1] 马加力[1] 施敏[1] 

机构地区：[1]上海交通大学医学院附属同仁医院消化内科,200336

出　　处：《肝脏》2015年第2期124-128,共5页Chinese Hepatology

基　　金：上海市医学重点学科建设基金资助(ZK2012A05);上海市卫生计生委科研课题资助(20134100)

摘　　要：目的探讨肝星状细胞通过CCL22/CCR4轴促进肝癌细胞侵袭的作用和可能机制。方法 TGF-β1(10 ng/ml)处理肝星状细胞LX-2不同时间,Western印迹检测处理前后肝星状细胞的活化标志物desmin及α-SMA的表达变化以及肝星状细胞LX-2和不同肝癌细胞系CCL22、CCR4表达。Transwell实验检测星状细胞LX-2或CCL22基因沉默后的MHCC-LM3侵袭的影响,Westem印迹检测上皮标志E-cadherin和间质标志N-cadherin及Vimentin的表达变化。结果肝星状细胞LX-2能被TGF-β1(10 ng/ml)活化,其活化的标志物desmin(24h:2.038±0.341;36h:2.562±0.248,相对于0h:1.329±0.172,P=0.008及P<0.001)和α-SMA(24h:2.741±0.439;36h:3.126±0.521,相对于0h:1.271±0.182,P<0.001)在处理24、36h后显著升高,星状细胞的趋化因子CCL22也随之高表达(24h:1.523±0.263;36h:1.836±0.319,相对于0h:3.126±0.634,P<0.001),3株人肝细胞癌细胞系均有CCR4的表达,其中MHCC-LM3细胞表达最强。肝星状细胞LX-2活化后与肝癌细胞MHCC-LM3共培养能诱导其上皮间质分化,活化24h和36h后上皮标志物Ecadherin表达明显下降(24h:1.273±0.207;36h:1.405±0.141,相对于0h:3.126±0.634,P<0.001),间质化指标Ncadherin(24h:2.516±0.384;36h:2.392±0.416,相对于0h:1.058±0.021,P<0.001)和Vimentin(24h:2.875±0.437;36h:2.924±0.353,相对于0h:1.452±0.121,P<0.001)表达则升高,并促进其侵袭。通过RNA干扰技术靶向沉默肝星状细胞CCL22则不能增强肝癌细胞侵袭能力,也不能诱导肝癌细胞MHCC-LM3发生上皮间质分化。结论肝星状细胞通过趋化因子CCL22/CCR4轴促进肝癌细胞侵袭,其机制可能与诱导肝癌上皮间质化有关。Objective To explore the impact of activated hepatic stellate cell on hepatocellular carcinoma invasion through CCL22/CCR4 axis.Methods The expressions of CCL22 and CCR4 were examined in hepatic stellate cell LX-2 and4 hepatocellular carcinoma cell lines by western blot at protein levels,respectively.In addition,Transwell invasion assay was carried out to analyze the influence of hepatic stellate cell LX-2 gene activating or CCL22 gene silencing on invasion of hepatocellular carcinoma cell MHCC-LM3.Western blot was performed to evaluate the expressions of epithelial marker Ecadherin and mesenchymal marker N-cadherin and Vimentin.Results Hepatic stellate cells LX-2 could be activated by TGF-β1（10 ng/ml）,and the expressions of desmin andα-SMA significantly increased in LX-2 with TGF-β1 stimulating for 24 h and 36 h,respectively.Meanwhile,chemokine CCL22 in LX-2 cells also rose.The expression of CCL22 was high in hepatic stellate cell LX-2,and expression of CCR4 was also found in all hepatocellular carcinoma cells.Hepatocellular carcinoma cells MHCC-LM3,which was co-cultured with hepatic stellate cell LX-2,induced the epithelial mesenchymal transition and increased the invasion.Furthermore,inhibition of CCL22 by gene silence in LX-2 suppressed the enhanced invasion and epithelial mesenchymal transition of MHCC-LM3 cells which was induced by activated hepatic stellate cells.Conclusion Hepatic stellate cells induce hepatocellular carcinoma cell invasion through chemokine CCL22/CCR4 axis,of which the mechanism might involve the epithelial mesenchymal transition of hepatocellular carcinoma cell.

关 键 词：肝星状细胞 肝癌 侵袭 上皮间质化 CCL22 CCR4 

分 类 号：R735.7[医药卫生—肿瘤]