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目的通过对MEL细胞在二甲基亚砜(DMSO)诱导分化前后和不同的铁状态,观察转铁蛋白受体(TfR)mRNA和铁螯合酶mRNA含量的变化。方法MEL细胞体外培养,加DMSO向红系诱导分化,同时以未诱导细胞为对照,应用Northernblot检测TfRmRNA和铁螯合酶mRNA含量,观察加入转铁蛋白、去铁胺或抗TfR单抗和Epo后的作用。结果和结论①未诱导的MEL细胞,随着细胞的增殖,TfRmRNA含量增加;诱导的MEL细胞,虽然随着细胞分化成熟而丧失增殖能力,但当存在血红素合成时,TfRmRNA含量也增高,且增高幅度较非诱导组高。②未诱导的MEL细胞,降低细胞内铁含量(培养液中加入去铁胺或抗TfR单抗),可刺激TfRmRNA表达,增加细胞内铁含量(培养液中加入转铁蛋白),则抑制TfRmRNA的表达;诱导的MEL细胞,相同条件的铁状态时,细胞内TfRmRNA水平发生类似的改变,但变化的程度较小。③当培养液中加入Epo刺激红系细胞分化时,促进TfRmRNA的表达。上述结果提示,在红系细胞,TfRmRNA的表达除受细胞内铁调节外,也受细胞内血红素合成的影响。④未诱导的MEL细胞,虽然细胞增生旺盛,但铁螯合酶mRNA?
OBJECTIVE: To investigate the changes of mRNA and mRNA of iron transporter (TfR) mRNA in MEL cells before and after differentiation in dimethyl sulfoxide (DMSO) and in different iron states. Methods The MEL cells were cultured in vitro and DMSO was added to the erythroid cells for differentiation. At the same time, untreated cells were used as control. The mRNA levels of TfR mRNA and iron chelatase were detected by Northern blotting. The effects of transferrin, desferrioxamine or anti-TfR monoclonal antibody and Epo After the role. RESULTS AND CONCLUSION ① The untreated MEL cells showed an increase in TfR mRNA levels as the cells proliferated. Although the induced MEL cells lost their ability to proliferate as the cells matured, the content of TfR mRNA increased in the presence of heme synthesis, and The increase rate was higher than non-induction group. (2) The untreated MEL cells, reducing the intracellular iron content (deferoxamine or anti-TfR monoclonal antibody in the culture medium) stimulated the expression of TfR mRNA and increased the intracellular iron content (transferrin added to the culture medium), then inhibited TfR mRNA The level of TfR mRNA in cells induced by MEL under the same conditions was similar, but the degree of change was smaller. ③ When adding Epo to culture medium to stimulate erythroid differentiation, promote TfR mRNA expression. These results suggest that, in erythroid cells, TfR mRNA expression is regulated by intracellular heme synthesis in addition to intracellular iron regulation. ④ Not induced MEL cells, although cell proliferation, but iron chelatase mRNA?