K562细胞红系诱导分化过程中血红蛋白基因与过氧化氢酶基因的表达变化  

Gene expression of hemoglobin and catalase during K562 cells differentiation

作　　者：剧锦哲[1,2,3] 马强[1,2] 贾小娥 李冯锐[2] 魏春华 苏燕[1,2] JU Jinzhe;MA Qiang;JIA Xiaoe;LI Fengrui;WEI Chunhua;SU Yan(Institute of Blood Conservation,Baotou Medical College,Baotou 014060,China;Preclinical and Forensic Medicine,Baotou Medical College;Central Laboratory,The First Affiliated Hospital of Baotou Medical College)

机构地区：[1]内蒙古科技大学包头医学院血液保护研究所,内蒙古包头014060 [2]内蒙古科技大学包头医学院基础医学与法医学院 [3]内蒙古科技大学包头医学院第一附属医院中心实验室

出　　处：《包头医学院学报》2018年第7期56-58,71,共4页Journal of Baotou Medical College

基　　金：国家自然科学基金(81160214);内蒙古自然科学基金(2016MS0801);包头市科技计划项目(2015C2006-16);包头医学院博士基金(BSJJ201615);包头医学院科研基金项目青苗计划(BYJJ-QM201645);内蒙古自治区研究生科研创新项目(S201510127Y01);内蒙古自治区卫生和计划生育委员会科研计划项目(201701090)

摘　　要：目的:研究K562细胞红系诱导分化过程中,血红蛋白α(hemoglobin subunit alpha-1,HBA-1)、血红蛋白ε(hemoglobin subunit epsilon-1,HBE-1)基因与过氧化氢酶(catalase,CAT)基因表达的变化。方法:利用氯化高铁红素(Hemin)诱导K562细胞构建红系诱导分化模型。利用联苯胺染色法和CCK-8法检测不同浓度Hemin作用K562细胞不同时间后,K562细胞分化阳性率及细胞的生长抑制率,确定后续实验Hemin的作用浓度。利用反转录实时荧光定量PCR检测K562细胞红系诱导分化过程中HBA-1、HBE-1、CAT基因mRNA表达水平的改变。结果:Hemin诱导K562细胞红系分化过程中,随Hemin诱导时间延长、浓度升高,细胞分化染色阳性率及细胞生长抑制率均升高(P<0.05)。40μmol/L Hemin诱导12 h、24 h、48 h后,与未诱导组相比,HBA-1、HBE-1基因mRNA表达量随时间延长逐渐增高(P<0.05);CAT基因mRNA表达在Hemin诱导48 h时增高(P<0.05)。结论:Hemin诱导K562细胞红系分化过程中,HBA-1 mRNA先于HBE-1 mRNA表达量增加,同时CAT基因mRNA表达量随之增高,说明CAT在红系分化早期可能起重要的抗氧化作用。Objective:To study the gene expression of hemoglobin subunit alpha-1(HBA-1),hemoglobin subunit epsilon-1(HBE-1)and catalase(CAT)during K562 cells erythroid differentiation.Methods:Hemin was used to induce K562 cells erythroid differentiation.Firstly,K562 cells were grouped by Hemin with different concentration and treated time.Then,benzidine staining method and CCK-8 were used to observe the positive benzidine staining rate and growth inhibition rate in different groups to determine the optimal conditions.Lastly,the reverse transcription quantification real-time PCR(RT-qPCR)was adopted to detect the expression of HBA-1,HBE-1 and CAT during differentiation.Results:During erythroid differentiation,the cellular positive staining rate and proliferation inhibition rate were increasing with the increase of treating concentration and time(P<0.05).After 12,24 and 48h of being induced by 40μmol/L hemin,the expression of HBA-1 and HBE-1 mRNA was increased gradually with the extension of time(P<0.05);the expression of CAT was increased at 48h(P<0.05).Conclusions:During K562 cells erythroid differentiation,the expression of HBA-1 mRNA increased before HBE-1.The increasing expression of CAT suggests that CAT may be regulated to be the essential antioxidase throughout the early differentiation.

关 键 词：K562细胞 红系诱导分化 血红蛋白 过氧化氢酶 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]