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目的探讨雷帕霉素(RAP)对急性淋巴细胞白血病细胞CEM-C1/C7糖皮质激素反应性的影响及可能的机制。方法以糖皮质激素抵抗株CEM-C1和糖皮质激素敏感株CEM-C7为研究对象,实验分为:1μmol/L地塞米松(DEX组);1μmol/L地塞米松+50 nmol/L雷帕霉素(DEX+RAP组);50 nmol/L雷帕霉素(RAP组);同时设对照组加入等体积无水乙醇。流式细胞仪检测细胞凋亡和细胞周期,CCK-8法检测细胞存活率,实时定量PCR检测CyclinD1 mRNA,Western blot检测CyclinD1蛋白水平。结果与其他组相比,DEX+RAP组可提高CEM-C1/C7的细胞凋亡率,降低细胞存活率,增加CEM-C1细胞周期阻滞在G0/G1期的比例(P<0.05);此外合用地塞米松和雷帕霉素可下调CyclinD1 mRNA和蛋白的表达。结论雷帕霉素通过增加细胞周期阻滞来提高急性淋巴细胞白血病细胞对糖皮质激素的敏感性。
Objective To investigate the effects of rapamycin (RAP) on CEM-C1 / C7 glucocorticoid responsiveness in acute lymphoblastic leukemia and its possible mechanism. Methods Corticosteroid-resistant CEM-C1 and glucocorticoid-sensitive CEM-C7 were used as experimental subjects. The experiment was divided into 1μmol / L dexamethasone (DEX group), 1μmol / L dexamethasone + 50 nmol / (DEX + RAP group) and rapamycin 50 nmol / L (RAP group). At the same time, the control group was given equal volume of absolute ethanol. Cell apoptosis and cell cycle were detected by flow cytometry. CyclinD1 mRNA was detected by CCK-8 assay and CyclinD1 protein by Western blot. Results Compared with other groups, DEX + RAP group could increase the apoptosis rate of CEM-C1 / C7 cells, decrease the cell survival rate and increase the proportion of CEM-C1 cell cycle arrest in G0 / G1 phase (P <0.05). In addition, combination of dexamethasone and rapamycin can down-regulate CyclinD1 mRNA and protein expression. Conclusion Rapamycin can increase glucocorticoid sensitivity of acute lymphoblastic leukemia cells by increasing cell cycle arrest.