THP-1细胞系白血病干细胞的检测、分选及鉴定  

Sorting and identification of leukemia stem cells in THP-1 cell line

作　　者：王颖超[1,2] 冯磊[1,2] 殷楚云 魏永纬[1,2] 马丽娜 王春美[1,2] 盛光耀 

机构地区：[1]郑州大学第一附属医院儿科 [2]河南省高等学校临床医学重点学科开放实验室,郑州450052

出　　处：《郑州大学学报（医学版）》2013年第2期210-213,共4页Journal of Zhengzhou University(Medical Sciences)

摘　　要：目的:探讨人急性单核细胞白血病THP-1细胞株中是否存在白血病干细胞(LSCs),并比较该LSCs与非LSCs亚群的生物学特性。方法:应用流式细胞术测定THP-1细胞中LSCs(CD34+CD38-细胞)的比例并分析LSCs与非LSCs的细胞周期。实时荧光定量PCR检测2种细胞中ABCG2、ABCB1耐药基因和Bcl-2、Bax凋亡基因的表达。将分选的LSCs移植到NOD/SCID小鼠,观察其致瘤能力。结果:THP-1细胞中存在CD34+CD38-的LSCs,比例为(0.12±0.06)%。大部分的LSCs处于细胞静止期,G0/G1期比例达(94.03±1.61)%;非LSCs增殖期细胞比例相对较高,S/G2/M期细胞比例为(56.58±1.59)%。实时荧光定量PCR结果显示:LSCs中ABCG2、ABCB1及Bcl-2的表达水平高于非LSCs(t=20.725、22.179、13.707,P<0.001),Bax在两亚群的表达无差异(t=1.635,P=0.071),但是LSCsBcl-2/Bax比值高于非LSCs(t=10.781,P<0.001)。1×103数量的LSCs可在4周后形成典型的NOD/SCID小鼠白血病浸润模型,未分选的THP-1细胞至少需要1×106才能成瘤。结论:THP-1细胞株中存在LSCs,这群细胞比例低,大部分处于静止期,具有干细胞生物学活性及体内致瘤性,可能与白血病的耐药和复发有关。Aim： To explore whether the human acute monocytic leukemia THP-1 cell line contains leukemia stemcells （ LSCs） or not,and to compare the biological properties between LSCs and non-LSCs. Methods： Flow cytometry was used to detect the percentage of LSCs （ CD34 ＋ CD38 -cell） in THP-1,and then the cell cycle of LSCs and non-LSCs was compared. The mRNA expressions of multidrug resistance proteins （ ABCG2,ABCB1） and apoptosis gene （ Bcl-2,Bax） were examined by real time quantitative PCR in two subpopulations. NOD / SCID mice were injected with different subpopu-lations of THP-1,and the incidence of leukemia was compared. Results： LSCs almost accounted for （ 0. 12 ± 0. 06） % of viable cells in THP-1,most of which were in rest stage,and the G 0 / G 1 phase cells accounted for （ 94. 03 ± 1. 61） % . While the S / G 2 / M phase cells in non-LSCs accounted for （ 56. 58 ± 1. 59） % . The mRNA expressions of ABCG2,ABCB1 and Bcl-2 in LSCs were higher than those in the non-LSCs （ t = 20. 725,22. 179,13. 707,P 0. 001） . Although there were no differences between these two subpopulations in Bax expression（ t = 1. 635,P = 0. 071） ; Bcl-2 / Bax value in LSCs was significantly higher than that in the non-LSCs （ t = 10. 781,P 0. 001） . The systemic disseminated leukemia model was es-tablished successfully by injecting 1 × 103 LSCs in 4 weeks after injection. Non-LSCs requires at least 1 × 106 to establish tumor model. Conclusions： The THP-1 cell line contains LSCs,and the proportion of LSCs is low. Most LSCs stay quies-cent and have biological property of stem cells as well as strong tumorigenic property,and they may be associated with tumor resistance and recurrence.

关 键 词：THP-1细胞 白血病干细胞 ABC转运体 NOD SCID小鼠 

分 类 号：R733[医药卫生—肿瘤]