人肝癌细胞系MHCC-97H干细胞体外培养及其生物学行为观察  

Culture of human liver cancer stem cells MHCC-97H and research of its biology behavior in vitro

作　　者：常军[1] 钱丽霞[1] 成岗[1] 吴清华[1] 顾琛[1] 姜建威[1] 

机构地区：[1]无锡市第三人民医院,江苏无锡214000

出　　处：《山东医药》2012年第39期39-41,共3页Shandong Medical Journal

基　　金：中华国际医学交流基金(编号CIMF-F-H001-114)

摘　　要：目的培养人肝癌细胞系MHCC-97H中具有干细胞特性的细胞群,探讨其分子生物学特点。方法将人肝癌细胞系MHCC-97H分别培养于干细胞培养体系及普通培养体系(分别为观察组及对照组)。培养10 d后显微镜(100倍及400倍)观察细胞生长情况;免疫荧光法检测细胞表面蛋白CD133、细胞核相关抗原(Ki67)、基质金属蛋白酶9(MMP9)、多药耐药蛋白1(MDR1)表达;West-blot法检测相应蛋白表达。结果观察组细胞呈半贴壁悬浮生长的克隆球;对照组细胞呈贴壁生长。观察组CD133、MDR1、MMP9呈高表达,对照组未见表达,P<0.05;两组Ki67均呈高表达。结论 MHCC-97H中存在一定数量的肿瘤干细胞,该群细胞是介导肝癌发生转移和耐药的主要因素。Objective To culture the human liver cancer lines MHCC-97H with stem cell characteristics in vivo, and to investigate its biology behavior. Methods Cultured the human liver cancer cells in serum-free medium and normal me- dium respectively（ observed group and control group respectively）. Ten days later, microscope was used to detect the cell growth state,immunofluorescence and West-blot assay were carried out to display the protein of CD133, MMP3, Ki67, MDR1 expression level. Results The cells in the observed group grown in the way of semi-adherent floating tumor spheres; while which in the control group grown in the way of adhesion. CD133 ,MDR1 ,MMP9 expressed highly in the ob- served group, but no expression were found in the control group（P 〈 0.05） ;Ki67 expressed highly in the two groups. Conclusion There exist a subpopulation of cells with characteristic of stem cell in human liver cancer cell MHCC-97H line, which maybe the main factor that mediated the metastasis and drug-resisitance of the hepatic carcinoma.

关 键 词：肝癌 肿瘤干细胞 细胞核相关抗原 基质金属蛋白酶3 多药耐药蛋白1 

分 类 号：R735.7[医药卫生—肿瘤]