全反式维甲酸诱导K562细胞分化过程中线粒体铁蛋白表达的研究  

Expression of Mitochondrial Ferritin in K562 Leukemic Cell During ATRA-induced Cell Differentiation

作　　者：孙蕾[1] 廖玍 高举[3] 陈婷婷[3] 潘琳丽[3] 袁粒星[4] 

机构地区：[1]四川大学华西第四医院内科,成都610041 [2]四川大学口腔疾病研究国家重点实验室,成都610041 [3]四川大学华西第二医院儿科,成都610041 [4]四川大学华西第二医院公共实验室,成都610041

出　　处：《四川大学学报（医学版）》2010年第1期77-80,90,共5页Journal of Sichuan University(Medical Sciences)

摘　　要：目的研究K562白血病细胞在全反式维甲酸(ATRA)诱导分化过程中线粒体铁蛋白(MtF)、运铁蛋白受体1(TfR1)和铁蛋白(Fn)mRNA表达水平的变化情况,探讨MtF在白血病细胞增殖和铁代谢方面的作用。方法K562细胞加入ATRA(终浓度1μmol/L)中诱导培养,分别于第1、3、5 d收集细胞,分别进行:①瑞士染色观察各时间点的细胞形态;流式细胞学检测细胞表面分化抗原CD13的表达;②Trizol法提取各时间点细胞的RNA,通过半定量RT-PCR方法检测各时点MtF、TfR1和Fn基因的表达水平。同时设未用ATRA诱导培养的K562细胞为对照组。结果1μmol/L的ATRA可诱导K562细胞向粒系细胞分化,诱导培养第5 d,细胞表面CD13的表达水平增加,诱导分化率为21.2%,与对照组比较,差异有统计学意义(P<0.05)。诱导分化前K562细胞即有MtFmRNA表达,但随细胞诱导时间的延长,MtF和TfR1 mRNA表达呈下降趋势,诱导分化第5 d的表达水平分别为诱导分化前的86.5%和79.2%;而FnmRNA的表达呈上调趋势,诱导分化第5 d表达水平为诱导分化前的1.21倍。结论ATRA诱导K562细胞向粒系细胞分化过程中,MtF和TfR1 mRNA表达下调,而FnmRNA表达上调,这种协调变化有助于降低细胞通过TfR1介导的铁摄取,从而抑制或影响细胞增殖潜能。Objective To investigate the expression of MtF, transferrin receptor 1 （TfR1） and ferritin （Fn） mRNAs in K562 leukemic cells during ATRA-induced cell differentiation and to explore the interrelationship between the expression levels of these iron metabolism-related molecules. Methods K562 cells cultured with or without ATRA （1 μmo]/L） were collected at 24, 72 and 120 hours respectively. Cell differentiation toward granulocyte lineage was confirmed by microscopic study （Wright＇s staining） and flowcytometry. Expression levels of MtF, TfR1 and Fn were evaluated with semiquantitative RT-PCR, while K562 cells cultured without ATRA as control. Results Over 21.2% of K562 cells demonstrated features of granulocyte, and the expression of CD13 on cell surface increased significantly at day 5 with ATRA treatment （P〈0.05, compared with control）. K562 cells could express a certain level of MtF before ATRA-induced differentiation. With increase of ATRA-induced cell differentiation, MtF mRNA expressions were downregulated progressively. After 5 days of induced cell differentiation, expression levels of MtF and TfR1 mRNA were just 86. 5% and 79. 2% of that before ATRA treatment. While Fn mRNA expression increased to 1.21 folds of that before ATRA treatment. Conclusion MtF expression is downregulated during ATRA-induced K562 cell differentiation, with concomitant downregulation of TfR1 and upregulation of Fn. The coordinated expression regulation of these key iron metabolism-related molecules during cell differentiation may in turn inhibit TfRl-mediated iron uptake via endocytosis and thus adversely affect cell proliferation potential.

关 键 词：ATRA K562细胞 线粒体铁蛋白 运铁蛋白受体1 铁蛋白 

分 类 号：R733.7[医药卫生—肿瘤]