白血病原代骨髓基质细胞对柔红霉素作用下Jurkat细胞基因表达的影响

Effects of leukemic bone marrow stromal cells on gene differential expression in human leukemic Jurkat T-cell line exposed to daurorubicin

目的研究白血病患者骨髓基质细胞(BMSCs)抑制柔红霉素(DNR)诱导 Jurkat 细胞凋亡过程中 Jurkat 细胞相关基因表达的变化,并进一步分析差异表达基因在 BMSCs 对 Jurkat 细胞保护过程中的意义。方法采用 Percoll 体外分离初治急性白血病患者骨髓单个核细胞,贴壁培养 BMSCs;采用抑制性消减杂交技术等分子生物学方法建立白血病 BMSCs 保护的 Jurkat 细胞差异表达基因的cDNA 文库;并对差异表达的基因进行初步鉴定与分析。结果成功地建立了白血病 BMSCs 保护的Jurkat 细胞上调和下调差异表达基因的 cDNA 文库;初步筛选克隆到30个上调差异表达基因和22个下调差异表达基因 cDNA 片段;其功能主要与细胞周期调控、细胞凋亡、细胞能量代谢相关。结论白血病患者 BMSCs 能诱导 Jurkat 细胞基因发生差异表达,为探讨白血病患者 BMSCs 保护白血病细胞逃避化疗药物杀伤的分子机制进行了前期研究。

Objective To investigate the inhibition effect of leukemic bone marrow stromal cells （BMSCs） on daunorubicin （DNR） induced apoptosis of human Jurkat cell line, and analyze the differentially expressed genes between Jurkat cells eocuhured with leukemic BMSCs or without. Methods Suppression subtractive hybridization （SSH） was employed to establish subtracted cDNA library of differentially expressed genes in Jurkat cells coeuhured with leukemic BMSCs and DNR. The cDNA fragments were sequenced and analyzed. Results The differentially expressed gene cDNA library was successfully developed. Primary screening was done by reverse Northern hybridization. Thirty up-regulated and 22 down-regulated eDNA fragments were isolated and sequenced. Analysis and comparison were performed in GenBank using BLAST. These genes are related to cell cycle regulation, cell apoptosis and energy metabolism. Conclusion Leukemic BMSCs influence gene expression of Jurkat cells. The resulting differentially expressed genes might be associated with the protection of leukemic cells by BMSCs from injury.