全反式维甲酸联合地西他滨对U937细胞系和初发老年AML患者的作用及机制研究  

All-Trans Retinoic Acid and Decitabine Synergistically Induce Anti-Leukemia Effect on U937 Cell Line and Newly Diagnosed Elder AML Patients

作　　者：董伟民[1] 曹阳[1] 向立丽[2] 林艳[1] 刘月 岑建农[3] 谢晓宝 顾伟英[1] DONG Wei-Min;CAO Yang;XIANG Li-Li;LIN Yan;LIU Yue;CEN Jian-Nong;XIE Xiao-Bao;GU Wei-Ying(Department of Hematology,The Third Affiliated Hospital of Suzhou University,The First People＇s Hospital of Changzhou,Changzhou 213003,Jiangsu Province,China;Department of Hematology,The Central Hospital of Xuzhou,Xuzhou 221009,Jiangsu Province,China;Jiangsu Institute of Hematology,The First Affiliated Hospital of Suzhou University,Suzhou 215006,Jiangsu Province,China)

机构地区：[1]苏州大学第三附属医院常州市第一人民医院血液科,江苏常州213003 [2]徐州市中心医院血液科,江苏徐州221006 [3]苏州大学第一附属医院江苏省血液病研究所白血病实验室,江苏苏州215006

出　　处：《中国实验血液学杂志》2018年第4期964-971,共8页Journal of Experimental Hematology

基　　金：江苏省333人才资助项目(BRA2015088);江苏省自然基金项目青年基金(BK20150253);常州市科技局应用基础项目(CJ20179052)

摘　　要：目的:分析全反式维甲酸(ATRA)联合地西他滨(DAC)对p16INK4a(p16)和维甲酸受体β(RARβ)的DNA甲基化以及基因表达的影响,探讨2药联合对U937细胞和老年初发急性骨髓系白血病(AML)患者的抗肿瘤作用及其机制。方法:采用实时荧光定量PCR和Western blot方法检测p16和RARβ的表达,通过甲基化特异性PCR分析p16和RARβ启动子的甲基化水平,应用WST-1法和流式细胞术分别检测ATRA联合DAC对U937细胞的增殖作用及其对分化、凋亡和细胞周期的影响。结果:ATRA联合DAC可以诱导DNA去甲基化,增强p16和RARβ的基因表达;2药联合导致U937细胞的生长抑制和分化、凋亡和周期阻滞;此外,对于不能耐受标准化疗的老年AML患者,ATRA联合DAC联合方案在发挥抗肿瘤活性的同时,伴随着p16和RARβ表达水平的上调,以及骨髓原始细胞的减少,且患者显示良好的耐受性。结论:ATRA联合DAC方案,作为诱导分化和去甲基化的联合治疗策略,具有抗AML作用潜能,有助于优化老年AML治疗策略和改善临床预后。Objective： To investigate the effect of all-trans retinoic acid（ ATRA） combined with decitabine（ 5-Aza-2＇-deoxycytidine; DAC） on DNA methylation and gene expression of p16 INK4 a（ p16） and retinoic acid receptor β（ RARβ）,and to explore their combined antineoplastic effect on U937 cells and newly diagnosed elder acute myeloid leukemia（ AML） patients. Methods： The expression levels of p16 and RARβ were determined by qRT-PCR and Western blot. Methylation-specific PCR was used to analyze their methylation status. WST-1 and flow cytometry were performed to detect growth inhibition,differentiation,apoptosis and cell cycle of U937 cells respectively. Results： The expression p16 and RARβ was down-regulated by promoter hypermethylation in newly diagnosed elder AML patients and U937 cells. Combination treatment of ATRA and DAC induced DNA hypomethylation as well as gene expression of p16 and RARβ,which contributed to the growth inhibition,differentiation,apoptosis and cell cycle arrest of U937 cells. In addition for elder AML patients intolerable to standard chemotherapy,the combination regimen of ATRA and DAC showed antineoplastic activity accompamied by up-regulation of p16 and RARβ expression and decrease of bone marrow blast,moreover the parients showed good tolerence to the reginen. Conclusion： The regimen of ATRA combined with DAC as the combination therapeutic strategy for inducing differentiation and demethylation possesses the anti-AML potency,and contributes to optimizing the therapeutic strategy for elder AML patients and promoting the clinical prognosis.

关 键 词：全反式维甲酸 地西他滨 急性髓性白血病 U937细胞 p16 RARβ 

分 类 号：R557.2[医药卫生—血液循环系统疾病] R733.7[医药卫生—内科学]