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

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-60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PT67并收获病毒上清,用病毒上清转染骨髓基质细胞系HFCL,经G418筛选得HFCL-pMSCV-Jagged1细胞,用Western印迹方法检测Jagged1基因的表达;将HL-60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-60细胞CD11b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-60细胞与HFCL-pMSCV-Jagged1、HFCL共培养48和72 h后,CD11b阳性率在Jagged1组明显低于HFCL组(P<0.05).结论:Jagged1基因表达可以较为显著地抑制ATRA诱导的HL-60细胞的分化.

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

Objective: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.