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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。
Objective To investigate the effect of leukemic bone marrow stromal cells on the cell cycle distribution of leukemia and its possible mechanism. Methods The primary normal and leukemic bone marrow stromal cells were isolated and cultured to simulate the function of bone marrow microenvironment and co-cultured with Jurkat cells to detect the cell cycle distribution of Jurkat cells. The suppression subtractive hybridization (SSH) screening and analysis of co-cultured leukemia bone marrow stromal cells Jurkat cells differentially expressed genes; RT-PCR technology to verify the expression of the target gene. RESULTS: The normal and leukemic bone marrow stromal cells were successfully isolated and cultured. After co-cultured with Jurkat cells for 24 h, the number of Jurkat cell cycle arrest (“G0 / G1” phase) was significantly increased (P <0.05) Hybridization assay showed that Jurkat cells up-regulated 30 genes and down-regulated 22 genes after 24 h co-culture with leukemia bone marrow stromal cells. RT-PCR results showed that Jurkat cells were highly expressed after 24 h co-culture with leukemia bone marrow stromal cells The CRIF1 gene is expressed. Conclusion Leukemia bone marrow stromal cells induce Jurkat cell cycle arrest (“G0 / G1” phase), which may be partially achieved by overexpression of CRIF1.