细胞自噬在儿童急性B淋巴细胞白血病化疗抵抗中的作用

Role of cellular autophagy in chemotherapy resistance of children with B-cell acute lymphocytic leukemia

目的以长春新碱(VCR)处理的人急性B淋巴细胞白血病细胞(BLCs)为模型,研究细胞自噬在儿童急性B淋巴细胞白血病(B-ALL)化疗中的作用。方法将BLCs随机分为对照组、药物组、阻断组和联合组,分别加入等体积的PBS、1μg/m L的VCR、10 mmol/L的细胞自噬抑制剂3-甲基腺嘌呤(3-MA)、1μg/m L的VCR+10mmol/L的3-MA处理48 h,采用WST-1法测算细胞增殖抑制率,免疫印迹法检测自噬相关蛋白轻链3β(LC3β)。结果处理细胞48 h后,药物组、阻断组和联合组细胞增殖抑制率分别为9.7%±3.3%、63.4%±9.1%、80.6%±8.3%;各组间两两比较,P均<0.01。对照组、药物组、阻断组和联合组LC3β蛋白表达量分别为0.02±0.01、0.31±0.05、0.01±0.01、0.02±0.01,药物组高于其他组(P均<0.01),其余各组间比较无统计学意义。结论细胞自噬可在ALL化疗过程中活化导致化疗抵抗。

Objective We take the human B-cell acute lymphocytic leukemia cells（ BLCs） treated with vincristine（ VCR） as the models to investigate the role of cellular autophagy in chemotherapy resistance of childhood B-cell acute lymphoblastic leukemia（ B-ALL）. Methods BLCs were randomly divided into the control group,experimental group,blocker group and union group,which were respectively treated with the same volume of PBS,1 μg / m L VCR,10 mmol / L 3-methyladenine（ 3-MA） and 1 μg / m L VCR ＋ 10 mmol / L 3-MA for 48 h. Transmission electron microscope was used to detect the autophagosome,WST-1 was employed to calculate the proliferation inhibitory rate and Western blotting was performed to detect the autophagy-related protein light chain 3β（ LC3β） expression. Results The inhibition rates after 48 h treatment in the experimental group,blocker group and union group were 9. 7% ± 3. 3%,63. 4% ± 9. 1% and 80. 6% ± 8. 3%,respectively（ all P 〈0. 01）. The LC3β expression in the control group,experimental group,blocker group and union group was 0. 02 ± 0. 01,0. 31 ± 0. 05,0. 01 ± 0. 01 and 0. 02 ± 0. 01,respectively,and the experimental group was higher than the other groups（ all P 〈0. 01）,no significant difference was found among the other three groups. Conclusion Autophagy can be activated during chemotherapy of ALL,leading to chemotherapy resistance.