去铁胺(DFO)诱导骨髓增生异常综合征(MDS)细胞株SKM-1的P15^(INK4B)基因去甲基化  

Defetoxamine(DFO)induces the demethylation of P15^(INK4B) gene in myelodysplastic syndrome(MDS)cell line SKM-1

作　　者：丁倩倩[1] 陈勤奋[1] 王小钦[1] 

机构地区：[1]复旦大学附属华山医院血液科,上海200040

出　　处：《复旦学报（医学版）》2014年第1期15-21,共7页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(81270583)~~

摘　　要：目的探讨铁螯合剂去铁胺(deferoxamine,DFO)诱导骨髓增生异常综合征(myelodysplastic syndrome,MDS)细胞株SKM-1的P15INK4B基因去甲基化作用。方法以枸橼酸铁铵(ferric ammonium citrate,FAC)和DFO处理SKM-1细胞,按不同铁负荷分成3组:对照组、FAC组和FAC+DFO组,分别检测不同铁负荷组细胞内可变铁池(labile ironpool,LIP)、细胞内活性氧类(reactive oxygen species,ROS)、P15INK4B基因甲基化状态、P15INK4B基因mRNA表达情况、细胞增殖(CFSE的平均荧光强度MFI)、细胞早期凋亡率以及细胞周期。结果与对照组相比,FAC组的LIP(64.04%±2.12%vs.1.45%±0.65%)、ROS(45.57%±1.18%vs.33.38%±12.96%)、细胞增殖MFI(23.01%±5.20%vs.51.67%±1.61%)明显升高,P15INK4B基因mRNA表达水平(0.72±0.08 vs.1)和细胞早期凋亡率(13.97%±2.25%vs.22.53%±1.76%)明显减少,差异均有统计学意义(P<0.05);与FAC组相比,FAC+DFO组的LIP(8.34%±4.21%vs.64.04%±2.12%)、ROS(34.39%±2.12%vs.45.57%±1.18%)、细胞增殖MFI(37.34%±6.61%vs.23.01%±5.20%)明显减少,P15INK4B基因mRNA表达水平(1.50±0.15 vs.0.72±0.08)和细胞早期凋亡率(55.07%±1.30%vs.13.97%±2.25%)明显升高,差异均有统计学意义(P<0.05);3组细胞周期的差异无统计学意义。结论 DFO可使LIP和ROS降低,诱导铁过载的SKM-1细胞P15INK4B基因去甲基化,使其mRNA重新表达,并可抑制铁过载的SKM-1细胞增殖,促进其凋亡。Objective To explore the demethylation effect of P15INK4B gene in myelodysplastic syndrome （MDS） cell line SKM-1 induced by iron chelating agents deferoxamine （DFO）. Methods SKM-1 cells were cultured with ferric ammonium citrate （FAC） ＋ DFO, and divided into three groups.. control group, FAC group and FAC ＋ DFO group. Cellular labile iron pool （LIP）, reactive oxygen species （ROS）, methylation state of P15mK4B gene, P15INK4B gene mRNA level, cellular proliferation （MFI of CFSE） ,cellular early apoptosis rate and cell cycle were detected. Results Compared with control group,the LIP （64.04% ± 2.12% vs. 1.45%＋ 0.65%）,ROS （45.57% ± 1.18% vs. 33.38% ±12.96%） and proliferation MFI （23. 01% ±5. 20% vs. 51. 67% ±1. 61%） were significantlyincreased,P15INK48 gene mRNA level （0.72 ±0.08 vs. 1） and early apoptosis rate （ 13.97% ＋ 2.25 % vs. 22.53% ＋ 1.76%） were significantly decreased in FAC group （P〈0. 05）. Compared with FAC group,LIP （8.34% ＋ 4.21% vs. 64.04% ± 2. 12%）,ROS （34.39% ＋ 2.12% vs. 45.57% ＋ 1.18%） and proliferation MFI （37.34% ± 6.61% vs. 23.01% ± 5.20%） were significantly decreased, P15INK4B gene mRNA level （1.50 ＋ 0. 15 vs. O. 72 ± 0.08） and early apoptosis rate （55.07% ± 1.30% vs. 13.97% ±2.25%） were significantly increased in FAC ＋ DFO group （P^0.05）. The cell cycle was not changed significantly. Conelusions Iron chelator agent DFO decreased LIP and ROS,induced the demethylation of P15INK4B gene, re-expressed P151nK4B gene mRNA, suppressed cellular proliferation, and increased cellular apoptosis in iron-overloaded SKM-1 cells.

关 键 词：骨髓增生异常综合征(MDS) SKM—1细胞 去铁胺(DFO) P15INK4B.基因 甲基化 

分 类 号：R551.3[医药卫生—血液循环系统疾病]