Xiang Lili, Dong Weimin, Wang Rong, Wei Jiang, Qiu Guoqiang, Cen Jiannong, Chen Zixing, Zheng Xiao, Hu Shaoyan, Xie Xiaobao, Cao Xiangshan, Gu Weiying
Department of Hematology, The First People's Hospital of Changzhou, Third Affiliated Hospital of Suzhou University, Changzhou, P.R. China.
Laboratory of China and United States Cooperation, The First People's Hospital of Changzhou, Third Affiliated Hospital of Suzhou University, Changzhou, P.R. China.
Oncol Lett. 2014 Jul;8(1):117-122. doi: 10.3892/ol.2014.2133. Epub 2014 May 12.
The aim of the current study was to investigate the antineoplastic activities of 5-aza-2'-deoxycytidine (also known as decitabine; DAC) and all- retinoic acid (ATRA), administered alone or in combination, in K562 cells , as well as the effects on the expression of the tumor suppressor genes, p16INK4a (p16) and retinoic acid receptor β (RAR-β). Cell growth inhibition, differentiation and apoptosis in K562 cells treated with DAC and/or ATRA were detected. The methylation of the p16 and RAR-β genes in the K562 cells was detected using the methylation-specific polymerase chain reaction (PCR) method. Quantitative PCR was used for the detection of the mRNA expression of the p16 and RAR-β genes, and western blot analysis was used to detect protein expression. DAC and ATRA, alone or in combination, had no effect on the growth inhibition, differentiation and apoptosis of the K562 cells. DAC alone induced the demethylation of the p16 gene, and combination of DAC and ATRA demonstrated more evident demethylation of the p16 gene, however, ATRA alone had no effect on methylation. The RAR-β promoter region was not methylated in the K562 cells. DAC in combination with ATRA appeared to produce a greater activation of the RAR-β gene, which led to the upregulation of the RAR-β expression level. ATRA enhanced the effect of DAC on p16 demethylation, and the combination of the two drugs was found to activate RAR-β expression, which indicated that DAC used in combination with ATRA has clinical potential in the treatment of human erythroleukemia.
本研究的目的是调查5-氮杂-2'-脱氧胞苷（也称为地西他滨；DAC）和全反式维甲酸（ATRA）单独或联合给药对K562细胞的抗肿瘤活性，以及对肿瘤抑制基因p16INK4a（p16）和维甲酸受体β（RAR-β）表达的影响。检测了用DAC和/或ATRA处理的K562细胞的细胞生长抑制、分化和凋亡情况。采用甲基化特异性聚合酶链反应（PCR）方法检测K562细胞中p16和RAR-β基因的甲基化情况。定量PCR用于检测p16和RAR-β基因的mRNA表达，蛋白质印迹分析用于检测蛋白质表达。DAC和ATRA单独或联合使用对K562细胞的生长抑制、分化和凋亡均无影响。单独使用DAC可诱导p16基因去甲基化，DAC与ATRA联合使用时p16基因去甲基化更明显，然而，单独使用ATRA对甲基化无影响。K562细胞中RAR-β启动子区域未发生甲基化。DAC与ATRA联合使用似乎能更大程度地激活RAR-β基因，并导致RAR-β表达水平上调。ATRA增强了DAC对p16去甲基化的作用，且发现两种药物联合使用可激活RAR-β表达，这表明DAC与ATRA联合使用在治疗人类红白血病方面具有临床潜力。
