复方中药制剂对急性髓系白血病细胞诱导分化与增殖的作用
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摘要
目的:研究复方中药制剂对白血病细胞的作用及其机制。方法:用复方中药制剂处理体外培养的髓系白血病细胞株,用CCK8的方法观察中药制剂对白血病细胞增殖的影响,瑞氏染色法观察细胞形态学的变化,流式细胞术检测细胞凋亡并测定细胞表面分化抗原CD11b的表达水平。结果:与单纯的4种白血病细胞系HL-60,MOLM-13,MV4-11,AML-M5相比较,不同中药浓度处理的这4种白血病细胞增殖能力减弱(P<0.05)且其增殖抑制作用呈剂量依赖性(r=0.9236;r=0.7488;r=0.8889;r=0.8119);与单纯的HL-60和AML-M5白血病细胞系相比,药物处理的这2种白血病细胞有明显的分化形态改变;与单纯的HL-60白血病细胞系相比,IC50中药浓度处理的HL-60细胞表面抗原CD11B增加85%±7.13%;与单纯的AML-M5白血病细胞系相比,1.5μl和2μl剂量中药制剂处理的AML-M5细胞的凋亡率增加(P <0.05)。结论:复方中药制剂对白血病细胞具有抑制增值作用,其机制可能与诱导白血病细胞分化和凋亡作用相关。
Objective: To study the effect of traditional chinese medicine (TCM) compound on myeloid leukemia cells and to explore its anti-leukemic mechanism. Methods: Myeloid leukemia cell lines were cultured in vitro and treated with TCM compound. The proliferation of the leukemia cells was measured by CCK8 method. The differentiation of the leukemia cells was evaluated by using Wright's staining method and by light microscopy, and the expression of differentiation-related surface antigens such as CD11B was measured and by flow cytometry, the apoptosis of the leukemia cells was detected by flow cytometry with using Annexin V staining. Results: Compared with untreated 4 leukemia cell lines HL-60, MOLM-13, MV4-11, AML-M5, the proliferations of 4 leukemia cells treated with different concentrations of TCM compound decreased (P < 0.05), and their proliferation inhibition were in a dose-dependent manner (r = 0.9236; r = 0.7488; r = 0.8889; r = 0.8119); compared with HL-60 and AML-M5 leukemia cells, the drugtreated 2 leukemia cells displayed obvious differentiated changes; compared with untreated HL-60 leukemia cell line,the expression of surface antigen CD11B increased by 85%±7.13% in HL-60 cells treated IC50 concentration of drug;compared with untreated AML-M5 leukemia cell line, the apoptotic rate of AML-M5 treated with 1.5 and 2 μl doses of TCM compound increased. (P < 0.05). Conclusion: The traditional chinese medicine compound may inhibit the proliferation of leukemia cell lines mainly by inducing leukemia cell differentiation and apoptosis.
Objective: To study the effect of traditional chinese medicine (TCM) compound on myeloid leukemia cells and to explore its anti-leukemic mechanism. Methods: Myeloid leukemia cell lines were cultured in vitro and treated with TCM compound. The proliferation of the leukemia cells was measured by CCK8 method. The differentiation of the leukemia cells was evaluated by using Wright's staining method and by light microscopy, and the expression of differentiation-related surface antigens such as CD11B was measured and by flow cytometry, the apoptosis of the leukemia cells was detected by flow cytometry with using Annexin V staining. Results: Compared with untreated 4 leukemia cell lines HL-60, MOLM-13, MV4-11, AML-M5, the proliferations of 4 leukemia cells treated with different concentrations of TCM compound decreased (P < 0.05), and their proliferation inhibition were in a dose-dependent manner (r = 0.9236; r = 0.7488; r = 0.8889; r = 0.8119); compared with HL-60 and AML-M5 leukemia cells, the drugtreated 2 leukemia cells displayed obvious differentiated changes; compared with untreated HL-60 leukemia cell line,the expression of surface antigen CD11B increased by 85%±7.13% in HL-60 cells treated IC50 concentration of drug;compared with untreated AML-M5 leukemia cell line, the apoptotic rate of AML-M5 treated with 1.5 and 2 μl doses of TCM compound increased. (P < 0.05). Conclusion: The traditional chinese medicine compound may inhibit the proliferation of leukemia cell lines mainly by inducing leukemia cell differentiation and apoptosis.
引文
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7李瑞芳,王庆端.冬凌草甲素对K562细胞端粒酶活性调控及细胞周期的影响.药学学报,2004;39(11):865-868.
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9汪丽佩,李天一,高瑞兰,等蟾蜍灵在体内外抑制急性红白血病细胞K562增殖及下调WT1表达的研究.中国药理学通报;2016;(2):229-233.
10邵淑丽,肖越,张伟伟,等.干扰MDR1基因表达提高急性早幼粒白血病HT9细胞对紫杉醇的敏感性.基因组学与应用生物学;2017;(4):1288-1293.
11张前杏,蔡文亮,颜平康,等.中医药治疗白血病研究进展.辽宁中医药大学学报,2015(9):110-113.
12 Burnett AK,Russell NH,Hills RK,et al.Arsenic trioxide and alltrans retinoic acid treatment for acute promyelocytic leukaemia in all risk groups(AML17):results of a randomised,controlled,phase3 trial.Lancet Oncol,2015;16(13):1295-1305.
13 Wang LL,Ren YQ.Efficacy analysis of arsenic trioxide combined with all trans retinoic acid for acute promyelocytic leukemia.中国实验血液学杂志,2015;23(5):1292-1295.
2 Becker PS,Medeiros BC,Stein AS,et al.G-CSF priming,clofarabine,and high dose cytarabine(GCLAC)for upfront treatment of acute myeloid leukemia,advanced myelodysplastic syndrome or advanced myeloproliferative neoplasm.Am J Hematol,2015;90(4):295-300.
3 Papaemmanuil E,Gerstung M,Bullinger L,et al.Genomic classification and prognosis in acute myeloid leukemia.N Engl JMed,2016;374(23):2209-2221.
4 Alex AA,Ganesan S,Palani HK,et al.Arsenic Trioxide Enhances the NK Cell Cytotoxicity Against Acute Promyelocytic Leukemia While Simultaneously Inhibiting Its Bio-Genesis.Front Immunol,2018;9(3):1357.
5孙启鑫,孟凡义,扶云碧,等.硼替佐米单用或联合三尖杉酯碱体外诱导HL-60细胞凋亡实验研究.中国实验血液学杂志,2007;15(2):233-236
6田艳伟.巴豆水提液对HL-60细胞的诱导分化作用.山西医药杂志,2002;31(3):265-265.
7李瑞芳,王庆端.冬凌草甲素对K562细胞端粒酶活性调控及细胞周期的影响.药学学报,2004;39(11):865-868.
8 Chen S J.A potential target of Tanshinone IIA for acute promyelocytic leukemia revealed by inverse docking and drug repurposing.Asian Pac J Cancer Prev,2014;15(10):4301-4305.
9汪丽佩,李天一,高瑞兰,等蟾蜍灵在体内外抑制急性红白血病细胞K562增殖及下调WT1表达的研究.中国药理学通报;2016;(2):229-233.
10邵淑丽,肖越,张伟伟,等.干扰MDR1基因表达提高急性早幼粒白血病HT9细胞对紫杉醇的敏感性.基因组学与应用生物学;2017;(4):1288-1293.
11张前杏,蔡文亮,颜平康,等.中医药治疗白血病研究进展.辽宁中医药大学学报,2015(9):110-113.
12 Burnett AK,Russell NH,Hills RK,et al.Arsenic trioxide and alltrans retinoic acid treatment for acute promyelocytic leukaemia in all risk groups(AML17):results of a randomised,controlled,phase3 trial.Lancet Oncol,2015;16(13):1295-1305.
13 Wang LL,Ren YQ.Efficacy analysis of arsenic trioxide combined with all trans retinoic acid for acute promyelocytic leukemia.中国实验血液学杂志,2015;23(5):1292-1295.