Atpif1基因对K562细胞血红蛋白合成的影响

Effects of Atpif1 gene on hemoglobin synthesis in K562 cells

目的:探讨线粒体三磷酸腺苷酶抑制因子-1(Atpif1)对血红蛋白合成的影响。方法:首先,将K562细胞分为低氧实验组、常氧对照组,低氧实验组采用O 2浓度为2%,分别培养24 h,48 h,72 h后收集两组细胞,通过细胞增殖-毒性检测试剂盒(CCK8)法检测细胞的活性,流式细胞仪检测低氧对细胞凋亡的影响,通过氯化血红素诱导K562细胞,检测低氧对血红蛋白合成的影响,qRT-PCR检测Atpif1,核因子(NF-κB),delta-氨基酮戊酸合成酶2(Alas2)基因的转录表达。然后,将K562细胞置于低氧培养箱培养并分为空白组,阴性对照组和si-Atpif1三组。转染siRNA,沉默Atpif1基因,观察血红蛋白合成和NF-κB、Alas2基因mRNA水平变化。结果:与常氧对照组相比,低氧实验组K562细胞活性降低、凋亡增加,血红蛋白含量增加(P<0.05)。Atpif1、Alas2、NF-κB的mRNA表达水平上调(P<0.05)。与空白对照组和阴性对照组相比,si-Atpif1组血红蛋白含量均有减少(P<0.05),同时NF-κB、Alas2的mRNA水平也出现下调(P<0.05)。结论:Atpif1基因参与调控血红蛋白合成,探究其在高原红细胞增多症(HAPC)发生中的作用,可以为防治HAPC提供新的思路和治疗靶点。

Objective:To investigate the effects of mitochondrial ATPase inhibitory factor 1(Atpif1)on hemoglobin synthesis.Methods:Firstly,the K562 cells were divided into 2 groups,hypoxia-treated group and normoxic control group.The K562 cells in hypoxia-treated group were treated with 2%oxygen.The K562 cells in the two groups were collected after cultured for 24,48 and 72 hours.The proliferation-inhibitory rates of cells were detected by CCK-8 assay.The apoptosis rates of K562 cells were analyzed by flow cytometry.The hemoglobin synthesis of K562 cells was induced by hemin.The gene expressions of Atpif1,Aladelta-aminolevulinate synthase 2(Alas2)and nuclear factor kappa B(NF-κB)were detected by qRT-PCR.Then,the K562 cells were cultured in hypoxic incubator and divided into blank control group,negative control group and si-Atpif1 group.After sliencing Atpif1 gene,the hemoglobin synthesis and the levels of NF-κB and Alas2 were determined.Results:Compared with the normoxic control group,the proliferation activity of K562 cells was inhibited,the apoptosis rate was increased,and the hemoglobin synthesis was also increased in hypoxia-treated groups.The expressions of Atpif1,Alas2 and NF-κB mRNA of K562 cells were upregulated.Compared with blank control group and negative control group,the content of hemoglobin was decreased,and the levels of NF-κB and Alas2 mRNA were also decreased in si-Atpif1 group.Conclusion:Atpif1 gene is involved in the regulation of hemoglobin synthesis.Exploring its roles in the development of high altitude polycythemia(HAPC)can provide new ideas and therapeutic targets for the prevention and treatment of HAPC.