白血病骨髓基质细胞黏附培养对Jurkat细胞bcl-2家族成员表达的影响

摘要

Objective To investigate the chemotherapeaties sensitivity and expression of bcl-2, bcl-xL and bax of Jurkat cells co-cultured with bone marrow stromal cells (BMSC) isolated and cultured from leukemia patients. Methods BMSC were isolated and cultured from leukemia patients routinely. To construct the co-cultured model, Jurkat cells were co-cultured with of the irradiated layer BMSC by 60Co and observed the model with scanning electron microscope. The Jurkat cells suspension-cultured were used as control. The apoptosis and IC50 were detected by the FACS can machine and MTT, respectively. The expression of bcl-2,bcl-xL and bax in Jurkat cells was detected by Western blotting. Results We found that the Jurkat cells in the model showed a decreased sensitivity to DNR, IC50 values for leukemic BMSC and nonadhered contol were of 2.30 μmol/L and 0.45 μmol/L, respectively. Moreover, Jurkat cells adhered to BMSC have a survival advantage over suspended cells following DNR exposure for 24 h, apoptosis percentages for leukemic BMSC group and nonadhered controls were of (6.05±0.54)% and (25.74±6.15)%, respectively. As compared with controls, leukemic BMSC group had significant difference in apoptosis percentages (P <0.01). The expression of bcl-2 in Jurkat cells was up-regnlated when adhered to BMSC for 4 h and the higher expression emerged after adhering for 24 h and 48 h. No marked change of bcl-xL and bax expressions were observed in the adhered Jurkat cells. Conclusion The adhered-culture with bone marrow stromal cells isolated from leukemia patients could make the leukemia cells acquire drug resistance, which was associated with the up-regulated expression of bcl-2 in the leukemia cells.%目的 观察白血病骨髓基质细胞(BMSC)-Jurkat细胞共培养模型中Jurkat细胞的化疗敏感性,并探讨基质细胞黏附培养对Jurkat细胞bcl-2、bcl-xL及box表达的影响.方法 常规分离、培养BMSC,Jurkat细胞与60Co照射的基质细胞层黏附培养,构建共培养模型,扫描电镜观察.以悬浮培养的Jutkat细胞为对照,MTT法测定Jurkat细胞DNR的IC50,TC-Annexin V/PI标记流式细胞仪定量Jurkat细胞的凋亡率,Western blotting检测Jurkat细胞总蛋白中bcl-2、bcl-xL和box的表达.结果 白血病基质细胞黏附使Jurkat细胞对DNR的敏感性降低,IC50卯为2.30μmol/L,而悬浮对照组为0.45μmol/L,白血病基质细胞黏附组Jurkat细胞凋亡率分别为(6.05±0.54)%,与悬浮对照组(25.74±6.15)%比较差异有统计学意义(P<0.01).Western blotting检测结果显示,与白血病BMSC黏附培养4 h即观察到bcl-2表达上调,24h和48h尤为明显,该家族另一凋亡抑制蛋白hcl-xL的表达未见明显变化,也未观察到促凋亡蛋白box表达的明显变化.结论 白血病BMSC黏附培养能介导白血病细胞耐药,其机制可能与基质细胞黏附使内血病细胞抗凋亡蛋白bcl-2表达上调有关.