Wu Di-jiong, Zhou Yu-hong, Zhu Jun, Zhao Wei, Zhong Wei-jun, Wang Zhen, Qian Huan, Li Rui, Fu Shan, Sun Jie
Hematology Department of TCM Hospital of Zhejiang Province, Hangzhou 310006, China.
Zhonghua Xue Ye Xue Za Zhi. 2011 May;32(5):313-6.
To explore the role of matrine (MAT) alleviating all-trans retinoic acid (ATRA) resistance in acute promyelocytic leukemia (APL) and its mechanism.
ATRA sensitive strain of APL (NB4) and resistant strain (NB4-R1, NB4-R2) were used in this study. The low toxic dosage of MAT was established by MTT test, and ATRA IC(50) of the cell strains (cultured with or without 0.1 mmol/L MAT) were obtained to confirm the reversal index (RI); the influence of MAT (10, 8, 6, 4, 2, 1, 0.1, 0.01, 0.001 mmol/L) combine with 1 µmol/L ATRA on the differentiation of the three cell strains were observed by nitro blue tetrazolium chloride (NBT) test and morphologic changes. The apoptosis rate of cells treated with different concentration of MAT combined with 1 µmol/L ATRA was tested by flow cytometry with Annexin V/PI staining.
(1) The toxicity of MAT to NB4, NB4-R1, and NB4-R2 cells was increased with the concentration, IC(50) value was (0.661 ± 0.035) mmol/L, (0.673 ± 0.132) mmol/L and (0.329 ± 0.020) mmol/L, respectively; (2) After treated with 0.1 mmol/L MAT, the ATRA resistance factor of NB4-R1 decreased markedly (RI = 4.96 ± 1.15), but did not of NB4-R2(RI = 0.66 ± 0.17); (3) The differentiation capacity of NB4 and NB4-R1 was enhanced with increase of MAT, and peaked at 0.1 mmol/L (P < 0.05), but did not of NB4-R2; (4) After treated with MAT, the ATRA (1 µmol/L) induced apoptosis rate in NB4 and NB4-R1 increased significantly (P < 0.05 and P < 0.01, respectively).
MAT can reverse the ATRA resistance of NB4-R1, which may relate to the effect of MAT on differentiation and apoptosis. Treatment with MAT plus ATRA may exaggerate the cells resistance potency.
探讨苦参碱（MAT）逆转急性早幼粒细胞白血病（APL）细胞全反式维甲酸（ATRA）耐药的作用及其机制。
采用APL的ATRA敏感株（NB4）及耐药株（NB4-R1、NB4-R2）。通过MTT实验确定MAT的低毒剂量，测定细胞株（加或不加0.1 mmol/L MAT培养）的ATRA半数抑制浓度（IC50）以确定逆转指数（RI）；通过氯化硝基四氮唑蓝（NBT）实验及形态学变化观察MAT（10、8、6、4、2、1、0.1、0.01、0.001 mmol/L）联合1 μmol/L ATRA对三株细胞分化的影响。采用Annexin V/PI染色，通过流式细胞术检测不同浓度MAT联合1 μmol/L ATRA处理后细胞的凋亡率。
（1）MAT对NB4、NB4-R1和NB4-R2细胞的毒性随浓度增加而增大，IC50值分别为（0.661±0.035）mmol/L、（0.673±0.132）mmol/L和（0.329±0.020）mmol/L；（2）用0.1 mmol/L MAT处理后，NB4-R1的ATRA耐药因子明显降低（RI = 4.96±1.15），而NB4-R2的未降低（RI = 0.66±0.17）；（3）随着MAT浓度增加，NB4和NB4-R1的分化能力增强，在0.1 mmol/L时达到峰值（P<0.05），而NB4-R2未增强；（4）用MAT处理后，ATRA（1 μmol/L）诱导的NB4和NB4-R1细胞凋亡率显著增加（分别为P<0.05和P<0.01）。
MAT可逆转NB4-R1的ATRA耐药，这可能与MAT对细胞分化和凋亡的作用有关。MAT联合ATRA治疗可能会增强细胞的耐药能力。
