罗丹明123分离肝癌细胞系HepG2异质性亚群的鉴定  

Isolation and identification of heterogenic subpopulation from the HepG2 cell line by Rhodamine123 extrusion assay

作　　者：千年松[1] 李韧[1] 于衡超[1] 曹云新[2] 张福琴[1] 窦科峰[1] 

机构地区：[1]中国人民解放军第四军医大学西京医院肝胆外科,陕西省西安市710033 [2]中国人民解放军第四军医大学基础医学部免疫学教研室,陕西省西安市710032

出　　处：《世界华人消化杂志》2008年第9期941-945,共5页World Chinese Journal of Digestology

摘　　要：目的:建立分离肝癌细胞系HepG2中对罗丹明123(Rho123)染料具有不同排斥能力细胞亚群的方法,并探讨其在肝癌异质性研究中的意义.方法:利用流式细胞分选术(FCM)并结合不同浓度的Rho123染料,分析和分选肝癌细胞系HepG2细胞中具有不同染料排斥能力的肝癌细胞亚群,再通过细胞生长的测定、软琼脂克隆形成以及免疫组织化学检测和裸鼠成瘤实验等方法,以比较不同肝癌细胞亚群的差异.结果:根据不同染料排斥能力可以将肝癌细胞系HeDG2细胞分为Rho^(low)和Rho^(high)2个亚群,2个亚群在倍增时间(16.9 h vs 24.7 h)、最大增生倍数(15.2 vs 12.3)、克隆形成率(50% vs 20%)以及甲胎蛋白(AFP)的表达(31/32 vs 20/26)以及裸鼠成瘤实验(7/10 vs 1/10)上具有显著性差异(均P<0.01),Rho^(low)亚群是具有干细胞特性的.结论:Rho123结合FCM可以富集具有干细胞特性的Rho^(low)亚群,同时进一步证实肝癌的异质性.AIM： To establish a method to separate the different subpopulations of HepG2 cell line according to the ability of these cells to extrude the Rho123 dye. METHODS： Using flow cytometry （FCM） in combination with different binding concentrations of Rho123 to analyze and sort subpopula- tions according to the ability of these cells to extrude the Rho123 dye. Viable cell count, soft agar cloning method, immunocytochemical staining and tumorigenicity investigation were performed to show the heterogeneity in Rho^low and Rho^high cells. RESULTS： Rho^low cells and Rho^high cells were separated using FACS analysis according to the ability of these cells to extrude the Rho123 dye. Growth rate, soft agar cloning（50% vs 20%）, expression of alpha-fetoproteins （AFP） （31/32 vs 20/26, P 〈 0.01） and tumorigenicity investigation （7/10 vs 1/10） were significantly increased in Rho123^low compared with those in Rho123h^high subpopulation. CONCLUSION： Subpopulations with stem cell properties of the HepG2 cell line can be enriched by Rho123/FCM, indicating that hepatocellular carcinoma is heterogeneous.

关 键 词：肝细胞癌 异质性 罗丹明123 肿瘤干细胞 流式细胞分选术 

分 类 号：R735.7[医药卫生—肿瘤] R737.14[医药卫生—临床医学]