表达人Jagged1基因的骨髓基质细胞系的建立及其在HL-60细胞分化过程中的作用  

Establishment of bone marrow stromal cell line expressing Jagged1 gene and its effects on differentiation of HL-60 cells

作　　者：宋献民[1] 袁有忠[1] 王健民[1] 杨建民[1] 江千里[1] 周虹[1] 

机构地区：[1]第二军医大学长海医院血液科,上海200433

出　　处：《第二军医大学学报》2003年第9期947-949,共3页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金 ( 3 9970 70 9)

摘　　要：目的 :建立表达人Jagged1基因的骨髓基质细胞系 ,并研究Jagged1基因表达在HL 6 0细胞诱导分化过程中的作用。 方法 :构建含有人全长Jagged1基因的逆转录病毒载体MSCV Jagged1 /neoR ,将其导入包装细胞PT6 7并收获病毒上清 ,用病毒上清转染骨髓基质细胞系HFCL ,经G4 1 8筛选得HFCL pMSCV Jagged1细胞 ,用Western印迹方法检测Jagged1基因的表达 ;将HL 6 0细胞与HFCL pMSCV Jagged1细胞、HFCL细胞在含有全反式维甲酸 (ATRA )的培养液中共培养 ,流式细胞仪检测不同时间点HL 6 0细胞CD1 1b的阳性率。 结果 :Western印迹方法证明HFCL pMSCV Jagged1细胞Jagged1表达水平明显高于HFCL细胞 ;HL 6 0细胞与HFCL pMSCV Jagged1、HFCL共培养 4 8和 72h后 ,CD1 1b阳性率在Jagged1组明显低于HFCL组 (P <0 .0 5 )。 结论 :Jagged1基因表达可以较为显著地抑制ATRA诱导的HL 6Objective:To establish the bone marrow cell line expressing human Jagged 1 gene and to observe its effects on the differentiation of HL 60 cells.Methods: Retro virus vector MSCV Jagged 1/neoR containing full length human Jagged 1 gene was constructed,and was used to infect the bone marrow stromal cell line HFCL.The cell line HFCL expressing human Jagged 1 gene was obtained with selection by G418 and detected with Western blot.HL 60 cells were co cultured with HFCL pMSCV Jagged 1 cells and HFCL cells respectively in the culture media containing all trans retinoic acid(ATRA).The cells expressing CD11b antigen were detected with flow cytometer at different time points after co culture.Results: Western blot showed the expression level of Jagged 1 gene in HFCL pMSCV Jagged 1 cells was higher than in HFCL cells.CD11b level was significantly less in Jagged 1 group than in HFCL group 48 h and 72 h after co culture( P <0.05).Conclusion: The expression of Jagged 1 gene can significantly inhibit the differentiation of HL 60 cells induced by the ATRA.

关 键 词：Jagged1基因 骨髓 基质细胞系 HL-60细胞 细胞分化 基因表达 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]