人胃癌侧群细胞的分离鉴定和生物学特性分析  

Isolation and characterization of side population cells in human gastric cancer cell line BGC-823

作　　者：吴婷[1] 李锦毅[2] 陆士新[3] 

机构地区：[1]辽宁医学院武警总医院研究生培养基地,北京100039 [2]武警总医院临床教研室 [3]中国医学科学院肿瘤医院肿瘤研究所病因室

出　　处：《中华肿瘤杂志》2012年第4期264-268,共5页Chinese Journal of Oncology

基　　金：国家自然科学基金，国家重点基础研究发展计划(973计划)项目

摘　　要：目的检测和分选胃癌细胞系中具有干细胞特性的侧群（sP）细胞，分析其生物学特性。方法制备胃癌细胞株BGC-823悬液，经Hoeehst33342染色后，用荧光激活细胞分选技术分选出人胃癌sP和非SP（NSP）细胞，测定sP细胞亚群所占的比例。通过平板克隆实验检测sP细胞的克隆形成能力，无血清培养检测sP细胞的增殖能力。以不同浓度的氟尿嘧啶（5-Fu）处理BGC．823细胞后，检测sP细胞的耐药性，通过小鼠体内成瘤实验检测sP细胞的致瘤性。结果流式细胞仪分析结果表明，BGC-823细胞中含少量sP细胞，占0．9％～2．1％。SP细胞组克隆形成数量为（217．67±15．50）个／孔，高于NSP细胞组[（147．00±17．58）个／孔，P〈0．01]；SP细胞组克隆形成率为（72．56±5．17）％，高于NSP细胞组[（49．00±5．86）％，P〈0．01]，表明sP细胞较NSP细胞有较强的克隆形成能力。经2．5、5、10和25斗∥ml的5-Fu作用36h后，sP细胞所占比例分别为12．9％、9．9％、4．0％和2．1％，化疗药物对sP细胞的抑制作用随着药物浓度的递增而加强。体内成瘤实验显示，sP细胞组瘤体质量为（0．176±0．034）g，NSP细胞组瘤体质量仅为（0．045±0．046）g，两组细胞所致瘤体质量差异有统计学意义（P〈0．05）。结论人胃癌细胞BGC-823中存在具有干细胞特性的sP细胞，进一彤开展对人冒痛千纲晌毕物学特忡的研容．将百r能棍示冒癌新的诊断和治疗涂释．Objective Isolate and characterize the side population （SP） cells with potency of stem cells from human gastric carcinoma cell line BGC-823. Methods SP and non-SP cells were sorted from BGC-823 cells by fluorescence-activated cell sorting （FACS） using Hoechst33342 staining. The tumorigenic ability of the SP cells was assessed by in vivo transplantation into non-obese diabetic/severe combined immunodeficiency mice. Results SP cells were isolated from BGC-823 cells in a proportion of 0.9% to 2.1% with respect to the whole cell population. The colony formation assay showed that the colony formation rate of the SP cells was significantly higher than that of the non-SP ceils （72.56% vs. 49.00%, P 〈 0.01 ）. The drug sensitivity test showed that the SP cells showed stronger drug resistance to 5-Fu than the non-SP cells. The in vivo transplantation of SP cells in mice showed that the tumor weight was （0. 176± 0.034）g, significantly higher thanthat of non-SP cells （0.045 ±0.046）g （P〈0.01）. Conclusions The results of this study indicate the existence of cancer stem-like SP cells in the human gastric cancer line BGC823 cells. Further characterization of this SP cell population may provide new insights for diagnosis and treatment of gastric cancer.

关 键 词：胃肿瘤 侧群细胞 肿瘤干细胞 流式细胞术 BGC-823细胞 小鼠 

分 类 号：R735.2[医药卫生—肿瘤]