Expression of the human growth hormone normal gene (hGH-N) in proliferating and differentiated HL-60 cells.
The possibility that human leukemic cells could synthesize growth hormone (GH) was investigated in the HL-60 cell line. Western blot analysis of protein extracts obtained from these cells revealed the existence of a major immunoreactive GH (irGH) band, with an approximate molecular weight of 22 kDa, together with lower amounts of 20- and 44-kDa bands. Stimulating proliferating HL-60 cells with KCl clearly increased GH concentration in the incubation medium as compared to basal values. RT-PCR amplification of HL-60 RNA and restriction assay of the amplimers demonstrated that those proteins were the result of the expression of the GH-N (normal) gene in this cell line. These results were confirmed by Northern blot, which also showed that the rate of GH-N gene expression was clearly dependent upon the proliferative state of the cells: while GH transcripts were easily detectable in actively proliferating cells, only minute amounts were observed when cells were induced to differentiate with dimethyl sulphoxide (DMSO). Similar differences were observed by Western blot. In all, these findings demonstrate that HL-60 cells are capable to produce and secrete a GH identical to pituitary GH. Interestingly, the rate of synthesis of the hormone dramatically increases when cells are actively proliferating. Therefore, it is likely that locally produced GH might be involved in the control of leukemic cell proliferation. Further studies are now in course to establish whether this mechanism occurs via an autocrine and/or paracrine way.
Department of Physiology, Faculty of Medicine, University of Santiago de Compostela, Spain.
J A
JA
Costoya
A
A
Vidal
M
M
García-Barros
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V
Arce
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J
Devesa
D001483:Base Sequence; D015153:Blotting, Western; D002454:Cell Differentiation / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D018922:HL-60 Cells; D019382:Human Growth Hormone / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics / Q000378:metabolism; D013004:Somatostatin / Q000378:metabolism
D017931:DNA Primers; D019382:Human Growth Hormone; D013004:Somatostatin; D012313:RNA