Ferrochelatase, glutathione peroxidase and transferrin receptor mRNA synthesis and levels in mouse erythroleukemia cells–mRNA synthesis and levels

Abstract

Mouse erythroleukemia Friend cells induced to undergo erythroid differentiation by treatment with hexamethylenebisacetamide (HMBA) were shown to increase cytoplasmic ferrochelatase mRNA, transferrin receptor (TfR) mRNA and glutathione peroxidase (GSHPx) mRNA levels. Inhibition of heme synthesis at the level of 5-aminolevulinic acid synthase by isonicotinic acid hydrazide (INH) and D,L-penicillamine (PA) or at the level of 5-aminolevulinic acid dehydratase by succinylacetone (SA) decreased the expression of ferrochelatase mRNA and TfR mRNA. In contrast with these mRNAs, the synthesis and the levels of glutathione peroxidase mRNA increased by the addition of these inhibitors of heme synthesis. The amount of iron in the intracellular low molecular mass iron pool detected in the post-mitochondrial supernatant of Friend cells treated with heme synthesis inhibitors was increased. On the other hand, iron levels in this pool declined with the preincubation of Friend cells with iron chelator pyridoxal isonicotinoylhydrazone (PIH). Further treatment with PIH or desferrioxamine (Desferal) increased the synthesis of TfR mRNA in induced Friend cells. The synthesis of ferrochelatase mRNA declined by the same treatment. The opposite was observed when the iron level in the low molecular mass intracellular nonheme iron pool was elevated by treatment with either diferric transferrin (Fe-Tf) or ferric pyridoxal isonicotinoylhydrazone (Fe-PIH). Exogenously supplied hemin stimulated the synthesis of ferrochelatase mRNA in uninduced Friend cells, while the synthesis of this mRNA in Friend cells taken on the fifth day after induction was inhibited by the addition of hemin.

Erythroid differentiation and protoporphyrin IX down-regulate frataxin expression in Friend cells: characterization of frataxin expression compared to molecules involved in iron metabolism and hemoglobinization

“…On erythroid differentiation there is a marked up-regulation of proteins involved in controlling Fe uptake (eg, TfR), 41,61 heme synthesis, 26,40,56,63 and hemoglobinization (eg, ␤-globin). 37,38 The marked up-regulation of TfR expression results in a significant increase in Fe uptake ( Figure 1B) as demonstrated by others.…”

“…In this study we used a well-characterized model of erythroid differentiation where incubation of Friend cells with DMSO results in initiation of heme biosynthesis, a significant increase in Fe uptake from Tf, and incorporation of Fe ϩϩ into PIX to form heme ( Figure 1B). 26,37,40,41,50,56,61,[63][64][65] Hence, our experiments have focused on the role of frataxin during a physiologic response rather than a pathologic condition.…”

“…26,60 Furthermore, although frataxin levels decreased during differentiation, there is increased expression of a variety of molecules involved in heme synthesis and other functions in the cytoplasm or mitochondrion. 26,[61][62][63][64][65] Indeed, there are only a few specific markers that are known to be down-regulated by DMSO treatment of Friend cells. In this respect, the decreases in Frda and Nramp2 gene expression are interesting (Figure 6), but they are not the only molecules that decrease during erythroid differentiation (eg, ribosomal proteins).…”

Mechanism of cell death by 5‐aminolevulinic acid‐based photodynamic action and its enhancement by ferrochelatase inhibitors in human histiocytic lymphoma cell line U937

“…It is well known that iron uptake in cells is mediated by the TfR, and that the intracellular iron level regulates cellular proliferation and heme synthesis. [38][39][40] Thus, the possible involvement of TfR in ALA-PDA was studied. Western blot analysis showed the presence of TfR in U937 cells as well as in different cancer cells, such as 253J-BV and PC12 cells (Figure 11(D)).…”

Transcriptomic and proteomic analysis of mouse radiation-induced acute myeloid leukaemia (AML)

“…Integrin Beta 3 (Itgb3) is essential for leukaemogenesis but dispensable for normal haematopoiesis hence suggesting that Itgb3 signalling pathway is a potential therapeutic target in AML [16]. Succinylacetone (SA; 4,6-dioxoheptanoic acid), a specific inhibitor of delta-aminolevulinic acid dehydrase (ALAD), leading to growth inhibition of leukaemia cells, [17] is associated with an important pathway in mouse erythroleukaemia cells [18]. In mouse, erythroleukaemias are associated with Friend virus infection where a clonal leukaemia develops through the proviral insertional activation of Spi1/Pu.1 leading to overexpression of PU.1 [19, 20].…”