摘要
目的观察白血病骨髓基质细胞-Jurkat细胞共培养模型中Jurkat细胞的化疗敏感性，探讨基质细胞黏附对Jurkat细胞周期及其调控蛋白表达的影响。方法常规分离、培养骨髓基质细胞，Jurkat细胞与^60Co照射的基质细胞层黏附培养，构建基质细胞-Jurkat细胞共培养模型，扫描电镜观察，通过MTT法测定Jurkat细胞DNR的IC50，流式细胞仪检测Jurkat细胞周期分布，Westernblot检测黏附培养4、24和48h Jurkat细胞周期调控蛋白cyclin A、cyclin E及p27的表达。结果悬浮培养Jurkat细胞的IC50为0．45μmol／L，而白血病基质细胞及正常基质细胞黏附组IC50分别为2．30μmol／L和1．78μmol／L。Jurkat细胞与基质细胞黏附培养24h，Go—G1期细胞的比例明显升高（48．74％±8．77％），与悬浮对照组（27．83％±1．86％）比较差异有统计学意义P〈0．01，G2-M期细胞比例明显降低为2．01％±1．17％，与悬浮对照组（20．33％±1．84％）比较差异有统计学意义P〈0．01；Western Blot检测发现cyclin A、cyclin E的表达在Jurkat细胞黏附培养4h时即有所下调，24h和48h尤为明显，而p27的表达则出现上调，以48h最为明显。结论白血病基质细胞黏附能介导白血病细胞耐药，其机制可能与白血病细胞周期改变有关。
Objective To investigate the effects of adhesion mediated by bone marrow stmma cells from leukemia patient on chemotherapeutics sensitivity and cell cycle of Jurkat ceils in the co-cultured model. Methods Bone marrow stroma cells were isolated and cultured from leukemia patients routinely. To construct the co-cultured model, Jurkat cells were co-cultured with BMSCs the irradiated layer by 60Co, and the model was observed with scanning electron microscope. The IC50 values of Jurkat cells exposured to DNR were quantified by MTT. The cell cycles of Jurkat cells after 24h-adhesion in the co-cultured model were analyzed by FACS. The expression of cyclin A, cyclin E and p27 in Jurkat cells adhered to BMSCs for 4h, 24h and 48h were detected by Western blot. Results Jurkat cells in the co-cultured model showed a decreased sensitivity to DNR, IC50 values for normal BMSCs, leukemic BMSCs and non -adhered control were of 1. 78μmol/L,2. 30μmol/L and 0. 45μmoL/L, respectively. The percentages of G0-G1 phase for leukemic BMSCs group and non - adhered control group were of 48.74% ± 8.77% and 27.83% ±1.86% , respectively. The percentages of G2-M phase for leukemic BMSCs group and non -adhered control group were of 2. 01% ±1.17% and 20. 33% ±1.84%, respectively. Compared with control group, the 24h-adhesion mediated by BMSCs from leukemia patients up-regulated the percentage of G0-G1 phase of Jurkat cells （ P 〈0. 01 ）, but down-regulated the percentage of G2-M phase （ P 〈 0. 01 ）. The expression of cyclin A and cyclin E in Jurkat cells was up-regulated when adhered to BMSCs for four hours and the higher expression emerged after adhering for 24h and 48h. Oppositely, the expression of p27 were down-regulated, especially after 48h-adhesion. Conclusions Drug resistance of leukemia cells can be mediated by adhering to bone marrow stroma cells, which may be related to the changes of cell cycle.
出处
《中国医师杂志》
CAS
2009年第1期50-53,共4页
Journal of Chinese Physician
