CAJ | 학술논문

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1 μmol/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 1 μmol/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MtF mRNA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Fn mRNA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Fn mRNA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.
목적 연구K562백혈병세포재전반식유갑산(ATRA)유도분화과정중선립체철단백(MtF)、운철단백수체1(TfR1)화철단백(Fn)mRNA표체수평적변화정황,탐토MtF재백혈병세포증식화철대사방면적작용.방법 K562세포가입ATRA(종농도1 μmol/L)중유도배양,분별우제1、3、5 d수집세포,분별진행:①서사염색관찰각시간점적세포형태;류식세포학검측세포표면분화항원CD13적표체;②Trizol법제취각시간점세포적RNA,통과반정량RT-PCR방법 검측각시점MtF、TfR1화Fn기인적표체수평.동시설미용ATRA유도배양적K562세포위대조조.결과 1 μmol/L적ATRA가유도K562세포향립계세포분화,유도배양제5 d,세포표면CD13적표체수평증가,유도분화솔위21.2%,여대조조비교,차이유통계학의의(P＜0.05).유도분화전K562세포즉유MtF mRNA표체,단수세포유도시간적연장,MtF화TfR1 mRNA표체정하강추세,유도분화제5 d적표체수평분별위유도분화전적86.5%화79.2%;이Fn mRNA적표체정상조추세,유도분화제5 d표체수평위유도분화전적1.21배.결론 ATRA유도K562세포향립계세포분화과정중,MtF화TfR1 mRNA표체하조,이Fn mRNA표체상조,저충협조변화유조우강저세포통과TfR1개도적철섭취,종이억제혹영향세포증식잠능.
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 μmol/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 TfR1-mediated iron uptake via endocytosis and thus adversely affect cell proliferation potential.