地西他滨对同步化的G0/G1期MDS-L细胞系线粒体功能的影响  

Role of decitabine in the mitochondria biogenesis of synchronized G0/G1 phase MDS-L cell line

作　　者：邱宗建 杨娟[1] 宋强[1] 

机构地区：[1]山东大学齐鲁医院血液科,山东济南250012

出　　处：《山东大学学报（医学版）》2014年第5期5-9,共5页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(Y2006C63)

摘　　要：目的初步探讨地西他滨(DAC)在不整合入基因组DNA的情况下对线粒体生物学功能的影响。方法采用阿非迪霉素(APC)将MDS-L细胞同步化在G0/G1期。DCFH-DA检测不同浓度DAC(0、5、10、15μmol/L)处理后的同步化G0/G1期细胞系活性氧(ROS)的产生水平。采用qRT-PCR法检测线粒体DNA拷贝数和线粒体编码基因NADH脱氢酶1、6(ND1、ND6)mRNA改变。结果与对照组相比,DAC在低浓度5μmol/L条件下可以通过不依赖DNA损伤的途径增加细胞ROS的产生(P<0.05),同时细胞内线粒体DNA(MtDNA)的拷贝数较对照组明显增加,差异有统计学意义(P<0.05)。但随着DAC浓度增加,DAC在高浓度(15μmol/L)可降低ROS的产生(P<0.05)。此外,在高浓度DAC作用下,ND1和ND6基因表达较对照组明显升高(P<0.05)。结论 DAC通过影响细胞ROS产生,改变线粒体DNA的拷贝数以及调控线粒体基因的表达,产生对线粒体功能的影响,并呈浓度依赖性。Objective To investigate the effect of decitabin（DAC） on the mitochondrial biogenesis of synchronized G0/G1 cells. Methods MDS-L cells were treated with aphidicolin（APC） to synchronize cell cycle at G0/G1 stage. After treatment of decitabine at different concentrations（0,5,10,15 μmol/L）,reactive oxygen species（ROS） productions were detected by DCFH-DA. Changes of mitochondrial DNA copy number and expressions of coded genes, NADH dehydrogenase 1（ND1） and NADH dehydrogenase 6（ND6）,were detected with qRT-PCR. Results In comparison with the control group,decitabine at lowconcentration（5 μmol/L） could promote the production of ROS（P ＆lt; 0. 05） and increase the copy number of mtDNA（P ＆lt; 0. 05）; however,as the concentration increased to 15 μmol/L, ROS production started to decline to even lower than that of the control（P ＆lt; 0. 05）. Besides,DAC could significantly change the expressions of ND1 and ND6 at high concentration. Conclusion Decitabine can affect mitochondrial biogenesis by altering the mtDNA copy number and gene expressions in a concentration-dependent manner.

关 键 词：地西他滨 G0G1期 线粒体 活性氧离子 

分 类 号：R551.3[医药卫生—血液循环系统疾病]