PARP-1沉默对1,4-苯醌致骨髓间充质干细胞微核形成的影响  

Effect of Poly(ADP-ribose) Polymerase-1 Silencing on Micronucleus Induced by 1,4-benzoquinone in Bone Mesenchymal Stem Cells

作　　者：高羽亭 杨慧 

机构地区：[1]广东医科大学公共卫生学院劳动卫生与环境卫生教研室,东莞523808 [2]东莞市环境医学重点实验室,东莞523808 [3]广东医科大学公共卫生学院试验中心,东莞523808

出　　处：《基因组学与应用生物学》2016年第11期2925-2929,共5页Genomics and Applied Biology

基　　金：广东省自然科学基金(No.2015A030310532);广东省医学科研基金(No.A2016246);广东医科大学科研基金(No.2XB13012)共同资助

摘　　要：为探讨聚腺苷二磷酸核糖聚合酶-1(poly(ADP-ribose)polymerase-1,PARP-1)对1,4-苯醌(1,4-benzoquinone,PBQ)诱导骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)微核形成的影响。我们以空载体BMSCs为对照组,以PARP-1沉默BMSCs为处理组,免疫印迹法检测PARP-1蛋白表达量。磷酸盐缓冲液(PBS)溶解PBQ,分别以0μmol/L、2.5μmol/L、5.0μmol/L、10.0μmol/L、20.0μmol/L、40.0μmol/L、80.0μmol/L、160.0μmol/L和320.0μmol/L PBQ染毒两组细胞,应用MTT法检测细胞活力。根据细胞活力检测结果选择合适染毒浓度后进行微核试验,实时荧光定量-聚合酶链反应检测PARP-1基因表达。结果显示,沉默细胞PARP-1的蛋白表达量较对照组的细胞((1.00±0.03)倍)下降了85%(p<0.05)。染毒24 h后,相同剂量下两组细胞的活力并无显著性差异。随着PBQ染毒剂量增加,两组细胞PARP-1表达水平、微核率和核异常率均明显增高(p<0.05),且相同剂量下PARP-1沉默组细胞比对照组细胞微核率及核异常率明显增高(p<0.05)。结果表明,PARP-1沉默BMSCs在PBQ作用下更易发生DNA损伤,且PARP-1沉默不利于DNA损伤的修复。In order to explore the effect ofpoly （ADP-ribose） polymerase-1 （PARP-1） silencing on micronucleus induced by 1,4-benzoquinone （PBQ） in bone mesenchymal stem cells （BMSCs）, we sorted BMSCs with PARP-1 silencing as the treated group, while BMSCs with empty vector were considered as the control group. The expression of PARP-1 protein was detected by immunoblotting. PBQ dissolved in PBS buffer at the concentrations of 0 μmol/L, 2.5 μmol/L, 5.0μmol/L, 10.0 μmol/L, 20.0 pumol/L, 40.0 μmol/L, 80.0 μmol/L, 160.0 μmol/L and 320.0 μmol/L were respectively given to both cells. The cell viability was detected with MTT. According to MTT assay, the PBQ concentration was chosen for the following study. The rate of micronucleus was tested. Expressions of PA RP-1 was tested by realtime fluorescence quantitive PCR. Compared with the control group, the expression of PARP-1 protein was reduced by 85% （p〈0.05）. The difference of viability in both cells was not significant after exposure of different doses of PBQ for 24 h. The expression of PARP-1 and the rate of micronucleus and dyskaryosis showed more increase with the increase of PBQ dose （p〈0.05）. And the rate of micronucleus and dyskaryosis in treated group changed more than the control group treated with the same dose （p〈0.05）. BMSCs with PARP-1 silencing occurred DNA damage more easily. And PARP-1 silencing was not conducive to repairing DNA damage.

关 键 词：聚腺苷酸二磷酸核糖转移酶-1 1 4-苯醌 骨髓间充质干细胞 微核 

分 类 号：R114[医药卫生—卫生毒理学]