摘要
目的:观察从人肝癌细胞系HepG2中分离的CD117+细胞生物学行为,探讨肝癌中CD117+细胞亚群的干细胞特性.方法:采用无血清悬浮培养法培养HepG2细胞,流式细胞术检测HepG2及球体细胞中CD117的表达比例.用流式细胞分选技术从HepG2成球细胞中分离CD117+的肿瘤细胞,进行无血清悬浮培养,观察其成球能力.CCK-8法观察CD117+细胞的增殖能力和顺铂对CD117+细胞的抑制率,计算IC50和耐药指数(RI).结果:HepG2细胞能在无血清培养基中存活、增殖并形成细胞球,成球率为6.21%±2.03%;流式细胞检测发现球体细胞中CD117+细胞的比例比HepG2细胞提升了9倍;CD117+细胞在无血清培养基中成球率和增殖能力均显著高于未分选细胞和CD117细胞;CD117+细胞在各浓度的顺铂作用下抑制率均较未分选细胞和CD117细胞明显降低,三者IC50分别为12.229μmol/L、7.970μmol/L和7.345μmol/L,CD117+细胞和未分选细胞耐药系数RI为1.165和1.076.结论:人肝癌细胞系HepG2中的CD117+细胞是具有肿瘤干细胞特性的细胞亚群,CD117可能是肝癌干细胞的候选表面分子标志物.
AIM：To isolate CD117＋ cells from human hepatocarcinoma cell line HepG2 and to observe their biological behavior and stem cell properties.METHODS：HepG2 cells were cultured in serum-free medium.Flow cytometry was used to detect the expression of CD117 in HepG2 cells and sphere cells.CD117＋ and CD117 cells were sorted by FACS and their ability of proliferation and self-renewal was observed in vitro.Chemosensitivity to cisplatin was determined by CCK-8 assay.The inhibitory rate of cells treated with cisplatin,the half maximal inhibitory concentration （IC 50） and resistance index （RI） were measured.RESULTS：HepG2 cells could survive,proliferate and form sphere cells in serum-free medium.The sphere-forming rate was 6.21%±2.03%.The percentage of CD117＋ cells population in sphere cells increased by 9 folds compared to HepG2 cells.The sphere-forming ability and proliferation ability of CD117＋ subpopulation in serumfree medium were significantly higher than unsorted cells and CD117 cells.The inhibitory rate of CD117＋ cells treated with cisplatin of different concentrations was lower than un-sorted cells and CD117 cells.The IC 50 were 12.229μmol/L in CD117＋ cells,7.970μmol/L in un-sorted cells,and 7.345μmol/L in CD117 cells,and the RI of CD117＋ cells and un-sorted cells were 1.165 and 1.076,respectively.CONCLUSION：CD117＋ cells are a subpopulation of HepG2 cells with stem cell properties.CD117 may be a candidate surface marker for liver cancer stem cells.
出处
《世界华人消化杂志》
CAS
北大核心
2011年第36期3649-3655,共7页
World Chinese Journal of Digestology
基金
国家重点基础研究发展计划(973计划)基金资助项目
No.2009CB521804
国家科技重大专项基金资助项目
No.2011ZX09102-010-02~~
