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目的 :探讨 HL- 6 0细胞经全反式维甲酸 (ATRA)作用后对化疗药物阿糖胞苷 (Ara- C)诱导凋亡敏感性的变化。方法 :应用光镜检查凋亡细胞形态 ,DNA电泳检查梯状条带及流式细胞仪检测细胞周期分布、凋亡细胞率和 bcl- 2蛋白表达的阳性细胞率和相对荧光强度 (MFI)。结果 :ATRA0 .3mg/ L 作用 HL- 6 0细胞 72 h后 ,S期细胞显著减少至 32 .9% (P<0 .0 5 ) ,G0 / G1 期细胞明显增加达 5 8.5 % (P<0 .0 5 ) ,bcl- 2阳性细胞率和 MFI分别下降至 18%和 0 .6 3(P<0 .0 5 ) ;Ara- C1.5 m g/ L 作用 HL- 6 0细胞 4h,凋亡细胞率为 5 5 .1% ,DNA电泳见明显的梯状条带。当 HL- 6 0细胞经 ATRA0 .3m g/ L 作用 72 h后再加 Ara- C1.5 m g/ L 继续培养 4h,细胞凋亡率明显减少至 34 .4% (P<0 .0 5 ) ,DNA电泳见梯状条带亮度减弱。结论 :ATRA降低 HL- 6 0细胞对 Ara- C诱导凋亡敏感性 ,其机制可能与 ATRA阻滞 G0 / G1 期细胞进入 S期有关
Objective: To investigate the change of sensitivity of HL-60 cells to Ara-C induction after all-trans retinoic acid (ATRA) treatment. Methods: The morphology of apoptotic cells was examined by light microscopy. The cell cycle distribution, the percentage of apoptotic cells and the positive rate of bcl-2 protein and relative fluorescence intensity (MFI) were detected by DNA ladder and flow cytometry. Results: After treated with ATRA0.3mg / L HL-60 cells for 72 h, the number of S phase cells significantly decreased to 32.9% (P <0.05), and the percentage of cells in G0 / G1 phase increased significantly to 8.55% (P < 0.05). The percentage of bcl-2-positive cells and MFI decreased to 18% and 0.63 respectively (P <0.05), and HL-60 cells treated with Ara-C1.5 mg / The rate of dead cells was 51.5%, and DNA ladder showed obvious ladder-like bands. When HL-60 cells were treated with ATRA0.3 m g / L for 72 h and Ara-C1.5 mg / L was added for 4 h, the apoptosis rate was significantly decreased to 34.4% (P <0.05) , DNA ladder electrophoresis to see the brightness weakened. CONCLUSION: ATRA can reduce the sensitivity of HL-60 cells to Ara-C-induced apoptosis, and its mechanism may be related to the ATRA arrest of G0 / G1 phase cells into the S phase