中文摘要：

目的培养残留耐柔红霉素（DNR）的Jurkat细胞，并对其耐药机制进行初步探讨。方法体外分离培养初发急性淋巴细胞白血病患者骨髓基质细胞（BMSCs）以模拟骨髓造血微环境，在与Jurkat细胞长期共培养的过程中选用含50ng／ml DNR的培养液进行培养，随着培养时间的延长，绝大部分Jurkat细胞漂浮死亡。残留的Jurkat细胞逐渐恢复增殖活力，并逐渐获得抗药能力，可以在DNR终浓度为50ng／ml的培养液中存活、增殖，将该细胞记为Jurkat／DNR。绘制Jurkat细胞和Jurkat／DNR细胞的增殖曲线，并检测其细胞周期分布、对多种化疗药物的耐药系数、DNR摄药量以及MDR1、bcl-2、bax和MRP mRNA的表达。结果嵌合于基质细胞层中的Jurkat细胞在DNR的持续刺激作用下逐步产生耐药性，获得残留耐DNR的Jurkat细胞。与Jurkat细胞相比，Jurkat／DNR细胞中处于静止期的细胞比例增高、增殖速度缓慢、对多种化疗药物产生不同程度的耐药性、DNR摄药量减少，可以检测到MDR1基因表达，bax基因表达无显著差异，bcl-2及MRP基因表达水平升高。结论骨髓基质细胞滋养层是残留白血病细胞生存和再生长的微环境，可增强白血病细胞的抗药能力；通过改造白血病患者造血微环境有可能抑制残留白血病的形成及减少复发。

英文摘要：

Objective To cultivate the Jurkat cells with residual daunorubicirrresistance （DNR）, and primarily explore the mechanism of drug resistance. Methods To simulate the hematopoietic mi t in leukemia bone marrow, the bone marrow stromal cells （BMSCs） were isolated from the bone marrow of patients with acute leukemia and cultivated in vitro, and co-cultivated with Jurkat cells using daunorubicin （50ng/ml） enriched solution, continuously with the concentration increasing so as to screen the DNR Jurkat cells. The proliferation of DNR Jurkat cells, the cell cycle, the intake dose of daunorubicin, the resistant indexes to different drugs and the expressions of MDR1, bcl-2, bax and MRP mRNA were detected. Results With the continuous stimulation of daunorubicin, the Jurkat cells which imbedded in strornal cells layer survived and developed the drug resistant character. The percentage of DNR Jurkat cells in resting phase ceils was higher compared with that of Jurkat cells, while the velocity of proliferation and the intake dose of daunorubicin were lower. The expression of MDR1 mRNA was detected and the expressions of bcl-2 and MRP mRNA in DNR Jurkat cells were higher than that in Jurkat cells, while the expression of bax mRNA showed no changes. Conclusion Feeder layer of stromal cells is the microenviroment in which the survival leukemia cells live and regrow, and it can enhance the drug resistance of leukemia cells. It is implied that reconstruction of the leukemia hematopoietic microenviroment may be an efficient way to inhibit the survival leukemia and decrease the risk of relapse.