HCAR2介导低氧促K562细胞向红系分化作用  

HCAR2 mediates hypoxia-promoted erythroid differentiation of K562 cells

作　　者：陈迎 李建 曹炎 何云凌 吴丽颖 周钢桥 Chen Ying;L Jan;Cao Yan;He Yunling;Wu Liying;Zhou Cangiao(School of Life Sciences,Anhui Medical University,Hefei 230032;National Center for Protein Sciences State Key Lab of Proteomics,Institute of Radiation Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Bejing 100850;Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850)

机构地区：[1]安徽医科大学生命科学学院,合肥230032 [2]军事科学院军事医学研究院辐射医学研究所,蛋白质组学国家重点实验室,国家蛋白质科学中心,北京100850 [3]军事科学院军事医学研究院微生物流行病研究所,北京100850

出　　处：《安徽医科大学学报》2023年第8期1249-1255,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81730055)。

摘　　要：目的研究低氧下羟基羧酸受体2(HCAR2)对人慢性髓系白血病细胞K562向红系细胞分化的影响。方法在含氯化血红素(hemin)的培养基中培养细胞,比较常氧/低氧条件下K562细胞向红系分化的能力,以及在此过程中HCAR2的表达情况。进而利用遗传学手段干预HCAR2表达,分别使其在K562细胞中敲低或过表达,通过联苯胺染色、RT-qPCR、Western blot及流式细胞术检测HCAR2在低氧促K562细胞向红系分化中的作用。结果K562细胞向红系分化1、2、3 d后,HCAR2的表达水平和K562细胞向红系分化的能力均随时间逐渐升高,并在低氧条件下得到增强;在K562细胞中敲低HCAR2,诱导分化后红细胞生成减少,且红系分化标记分子CD235a、γ-珠蛋白(γ-globin)的mRNA和蛋白表达均降低,红系细胞膜表面分子CD71和CD235a双阳性细胞比例下降;相反,在K562细胞中过表达HCAR2,与对照组相比,诱导分化后红细胞生成增加,且CD235a、γ-globin的mRNA和蛋白表达水平均升高,CD71/CD235a双阳性细胞比例也有所增加。结论HCAR2介导了低氧促进K562细胞向红系分化。Objective To explore the effect of hydroxy-carboxylic acid receptor 2(HCAR2)on erythroid differentiation of human chronic myeloid leukemia K562 cells under hypoxia.Methods The cells were cultured in medium containing hemin to compare the ability of K562 cells to differentiate into erythroid cells under normoxia/hypoxia conditions.Then the expression of HCAR2 was interfered with genetic methods to knockdown or overexpress it in K562 cells,respectively.Benzidine staining,RT-qPCR,Western blot and flow cytometry were used to detect the role of HCAR2 in the erythroid differentiation of K562 promoted by hypoxia.Results On day 1,2,3 after differentiation,the expression level of HCAR2 and the erythroid differentiation ability of K562 cells increased gradually with time,and hypoxia enhanced the above phenotypes compared with normoxia.In HCAR2-silenced K562 cells,erythroid cells was reduced and the expression of erythroid differentiation markers CD235a and-globin decreased.Simultaneously,the proportion of double-positive cells expressing erythroid cell surface molecules CD71 and CD235a was lower than that of the control group.On the contrary,the overexpression of HCAR2 in K562 cells increased erythroid cells generation as well as elevated the expression levels of CD235a and-globin.In addition,the proportion of CD71/CD235a double-positive cells also increased in HCAR2-overexpressed K562 cells.ConclusionHCAR2 mediates hypoxia in promoting the erythroid differentiation of K562 cells.

关 键 词：红系分化 低氧 HCAR2 K562细胞 

分 类 号：Q254[生物学—细胞生物学]