人肝癌细胞系MHCC-97H干细胞体外培养及其生物学行为观察

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 medium 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 observed 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.