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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 study the changes of Jurkat cell related gene expression in leukemic bone marrow stromal cells (BMSCs) induced by daunorubicin (DNR) -induced Jurkat cell apoptosis and to further analyze the significance of differentially expressed genes in Jurkat cell protection by BMSCs. Methods Percoll was used to separate bone marrow mononuclear cells from newly diagnosed acute leukemia patients in vitro and BMSCs were adherently cultured. The cDNA library of Jurkat cells differentially expressed in leukemia BMSCs was established by molecular biology methods such as suppression subtractive hybridization. The differentially expressed Gene identification and analysis of the initial. Results The cDNA library of up-regulated and down-regulated differentially expressed genes in Jurkat cells protected by leukemia BMSCs was successfully established. Thirty up-regulated differentially expressed genes and 22 down-regulated differentially expressed cDNA fragments were screened by preliminary screening. The functions were mainly related to cell cycle regulation, Apoptosis, cellular energy metabolism related. Conclusions BMSCs from patients with leukemia can induce differentially expressed genes in Jurkat cells. A preliminary study was carried out to explore the molecular mechanism of leukemia patients’ protection of leukemia cells against chemotherapeutic drug killing in patients with leukemia.