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目的探讨雷帕霉素是否可以逆转地塞米松(DEX)耐药的急性T淋巴细胞白血病(T-ALL)细胞株CEM-C1细胞的GC耐药性。方法以儿童T-ALL GC耐药株CEM-C1和GC敏感株CEM-C7为研究对象,MTT法检测不同浓度(1 000 ng/ml、100 ng/ml、10 ng/ml、1 ng/ml和0.1 ng/ml)雷帕霉素及其联合1μmol/L地塞米松(DEX)作用后对细胞增殖的影响;细胞离心甩片瑞氏染色法观察细胞形态学变化;Annexin V/PI染色法检测对细胞凋亡的影响;PI染色法检测细胞周期改变。结果①不同浓度雷帕霉素作用于CEM-C1和CEM-C7细胞,对细胞生长的抑制作用均呈时间和浓度依赖性,联合应用1μmol/L DEX后,这一生长抑制作用显著增强,24 h IC50约为100 ng/ml。②以终浓度100 ng/ml雷帕霉素作用于CEM-C1和CEM-C7细胞18、24及48 h,细胞形态学与对照相比均无明显变化;Annexin V-PI双染流式细胞术检测均未见明显凋亡;细胞周期测定发现两种细胞均阻滞于G1期,S期细胞比例均明显降低。③以终浓度100 ng/ml雷帕霉素联合1μmol/L DEX作用于CEM-C1细胞后,瑞氏染色发现CEM-C1细胞于24 h出现细胞核改变,48 h出现明显凋亡;Annexin V/PI双染细胞凋亡检测显示细胞凋亡率较单用雷帕霉素及DEX时显著增加,18 h时早期凋亡细胞开始增多,24 h时最为明显;细胞周期测定显示S期细胞比率较单用雷帕霉素时显著降低,而G1期细胞显著增多。结论雷帕霉素能够逆转CEM-C1细胞对GC的耐药性,其机制有待进一步深入研究。
Objective To investigate whether rapamycin can reverse the GC resistance of dexamethasone (DEX) -resistant acute lymphoblastic leukemia (T-ALL) cell line CEM-C1. Methods T-ALL GC-resistant children CEM-C1 and CEM-C7 were used as the research objects. MTT assay was used to detect the effect of different concentrations (1 000 ng / ml, 100 ng / ml, 10 ng / ml, 1 ng / ml And 0.1 ng / ml rapamycin combined with 1 μmol / L dexamethasone (DEX) on the cell proliferation. Cell morphological changes were observed by Wright staining with Annexin V / PI staining Detection of apoptosis on the impact; PI staining to detect cell cycle changes. Results ① The effect of different concentrations of rapamycin on CEM-C1 and CEM-C7 cells in a time and concentration-dependent manner inhibited the growth of the cells. When combined with 1 μmol / L DEX, the growth inhibition was significantly increased. 24 h IC50 is about 100 ng / ml. ② The morphological changes of cells in CEM-C1 and CEM-C7 treated with rapamycin at a final concentration of 100 ng / ml for 18, 24 and 48 h did not show any significant changes. Annexin V-PI double staining flow cytometry No significant apoptosis was found in the test. The cell cycle assay showed that both cells were arrested in G1 phase and the proportion of S phase cells were significantly decreased. ③ After treated with 100 ng / ml rapamycin and 1 μmol / L DEX in CEM-C1 cells at final concentration of 100 ng / ml, CEM-C1 cells showed nuclear changes at 24 h and significant apoptosis at 48 h; Wistar Annexin V / The apoptosis of PI double staining cells showed that the apoptotic rate was significantly higher than that of rapamycin and DEX alone, and the number of early apoptotic cells began to increase at 18 hours and was most obvious at 24 hours. The cell cycle assay showed that the ratio of S phase cells Rapamycin alone was significantly lower, while the G1 phase cells increased significantly. Conclusion Rapamycin can reverse the drug resistance of CEM-C1 cells to GC, and its mechanism needs to be further studied.