微环境诱导肝细胞癌多药耐药的形成及机制  

Formation of multidrug resistance of hepatocellular carcinoma induced by microenvironment and its mechanism

作　　者：朱虹[1] 陈孝平[1] 罗顺峰[1] 关剑[1] 张万广[1] 张必翔[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肝胆胰外科研究所,武汉430030

出　　处：《中华实验外科杂志》2005年第11期1315-1318,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30371395);卫生部2001~2003年临床学科重点资助项目(卫规财发[2001]321号)

摘　　要：目的探讨微环境在肝癌多药耐药(MDR)表型形成中的作用,并初探其作用机制,为逆转肝癌耐药提供有效的新的分子靶点。方法模拟肝癌体内生长的局部微环境,使HepG2细胞分别在缺氧、低糖的微环境下生长或稳定整合HBX基因。应用荧光定量聚合酶链反应(PCR)技术和Western blot技术分别检测这些局部微环境因素作用下的HepG2细胞内多药耐药相关基因mdr1、肺耐药相关蛋白基因(LRP)、多药耐药相关蛋白基因(MRP1)和缺氧诱导因子-1α(HIF-1α)的mRNA和蛋白水平的表达。同时运用免疫细胞化学技术检测肝癌耐药细胞株中HIF-1α蛋白的表达,以及检测转染了HIF-1α质粒的HepG2细胞中上述相关多药耐药基因的表达。结果在缺氧、低糖环境下生长或稳定整合了HBX基因的HepG2细胞均不同程度地高表达多药耐药相关基因和HIF-1α;在肝癌耐药细胞株中HIF-1α蛋白高表达;稳定转染了HIF-1α质粒的HepG2细胞显著高表达多药耐药相关基因。结论肝癌生长的微环境可通过核转录因子HIF-1α调控多药耐药相关基因的表达,从而诱导肝癌多药耐药表型的形成;HIF-1α有望成为逆转肝癌耐药的新的分子靶点。Objective To explore the role of microenvironment in formation of multidrug resistance （MDR） of hepatocellular carcinoma （HCC） and its mechanism and to find a new and effective molecular target on reversing MDR of HCC. Methods Local microenvironment of HCC growth in vivo was mimicked.The expression of of mdrl, MRP1, LRP and HIF-1α genes was detected at mRNA and protein level using real-time fluorescent quantitative PCR and Western-blot technique in HepG2 cells respectively exposed to hypoxia, low glucose or transfected by plasmid HBX/pcDNA3. HIF-1α protein in liver cancer cell line resistant to multidrug was investigated by immunohistochemistry. Also the expression of related multidrug resistance genes was detected in HepG2 cells transfected by plasmid HIF-1α/PCDNA3 .Results In HepG2 cells respectively exposed to hypoxia, low glucose or transfected by plasmid HBX/pcDNA3, the expression of related multidrug resistance genes and HIF-1α was increased to varying degrees. HIF-1α protein in liver cancer cell line resistant to multidrug was overexpressed. The expression of related multidrug resistance genes was remarkably upregulated in HepG2 cells transfected by plasmid HIF-1α/PCDNA3. Conclusion Microenvironment of HCC growth can modulate the expression of related multidrug resistance genes via HIF-1α, which induces the MDR phenotype of HCC. HIF-1α probably becomes a new and effective molecular target on reversing MDR of HCC.

关 键 词：癌 肝细胞 多药耐药 缺氧诱导因子1α 多药耐药(MDR) 局部微环境 肝细胞癌 HIF-1Α蛋白 缺氧诱导因子-1α 多药耐药相关蛋白基因 Westernblot技术 

分 类 号：R735.7[医药卫生—肿瘤]