CRIF1介导白血病骨髓基质细胞诱导Jurkat细胞周期阻滞的探讨

目的研究白血病骨髓基质细胞对白血病细胞周期分布的影响及其可能机制。方法分离培养原代正常和白血病骨髓基质细胞体外模拟骨髓微环境功能,与Jurkat细胞共培养后,检测Jurkat细胞周期分布情况;抑制性消减杂交技术筛选和分析与白血病骨髓基质细胞共培养24 h后Jurkat细胞差异表达基因;RT-PCR技术验证目的基因的表达水平。结果成功分离培养正常和白血病骨髓基质细胞,体外与Jurkat细胞共培养24 h后,Jurkat细胞周期阻滞("G0/G1"期)细胞数明显升高(P<0.05);抑制性消减杂交试验发现,与白血病骨髓基质细胞共培养24 h后Jurkat细胞上调表达30个基因克隆、下调表达22个基因克隆;RT-PCR验证发现,与白血病骨髓基质细胞共培养24 h后,Jurkat细胞高表达CRIF1基因。结论白血病骨髓基质细胞诱导Jurkat细胞周期阻滞("G0/G1"期),可能部分通过高表达CRIF1实现。

CRIF1 mediated cell cycle arrest of Jurkat cells induced by leukemia marrow stromal cells

Objective To investigate the influence of leukemic bone marrow stromal cells(BMSCs) on the cell cycle of leukemic cells and the possible mechanisms.Methord Normal BMSCs and leukemic BMSCs were isolated and cultured to simulate a hematopoietic microenvironment in vitro,and then cocultured with Jurkat cells to determine the effect on the cell cycle distribution.Suppression subtractive hybridization(SSH) was used to analyze the differential gene expression after co-culturing Jurkat cells with normal BMSCs and leukemic BMSCs.The expression of the target genes was verified by RT-PCR.Results Normal BMSCs and leukemic BMSCs were successfully isolated and cultured.After coculture with Jurtkat cells for 24h in vitro,we found that the cell cycle of Jurkat cells was arrested in the G0/G1 phase.Suppression subtractive hybridization study found that there were 30 up-regulated genes and 22 down-regulated genes in Jurkat cells after 24 h coculture with bone marrow stromal cells.RT-PCR validation found that there was increased expression of CRIF1 in Jurkat cells after 24 h coculture with bone marrow stromal cells Conclusion Primary cultured leukemic BMSCs could induce cell cycle arrest of Jurkat cells in the G0/G1 phase increased expression of CRIF1 by leukemic cells.