Metallothionein RNA levels in HL‐60 cells Effect of cadmium, differentiation, and protein kinase C activation

Hanke, Tomáš; Tyers, Michael; Harley, Calvin B.

doi:10.1016/0014-5793(88)81051-2

Cited by 6 publications

(6 citation statements)

References 28 publications

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“…A decrease of free zinc parallels the expression of surface markers CD11b and CD14 during 1,25D 3 -induced differentiation, and chelation by TPEN promotes the development of monocyte surface markers and function. Furthermore, unlike other studies where MT is upregulated in the early phase of differentiation, coincident with its nuclear translocation [15,19], we detected no changes in MT expression of differentiating HL-60, confirming a previous study [35].…”

Section: Discussionsupporting

confidence: 85%

Cellular zinc homeostasis is a regulator in monocyte differentiation of HL-60 cells by 1α,25-dihydroxyvitamin D3

Dubben

Hönscheid

Winkler

et al. 2010

Journal of Leukocyte Biology

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It was reported previously that zinc-deficient mice show impaired lymphopoiesis. At the same time, monocyte numbers in these animals are increased, indicating a negative impact of zinc on monocyte development. Here, we investigate the role of zinc homeostasis in the differentiation of myeloid precursors into monocytes. Reduced gene expression of several zinc transporters, predominantly from the Zip family, was observed during 1 alpha, 25-dihydroxyvitamin D(3) (1,25D(3))-induced differentiation of HL-60 cells. This was accompanied by a reduction of intracellular-free zinc, measured by FluoZin-3. Amplifying this reduction with the zinc chelator TPEN or zinc-depleted cell-culture medium enhanced 1,25D(3)-induced expression of monocytic surface markers CD11b and CD14 on HL-60, THP-1, and NB4 cells. In contrast, differentiation of NB4 cells to granulocytes was not zinc-sensitive, pointing toward a specific effect of zinc on monocyte differentiation. Further, monocyte functions, such as TNF-alpha secretion, phagocytosis, and oxidative burst, were also augmented by differentiation in the presence of TPEN. The second messenger cAMP promotes monocyte differentiation. We could show that zinc inhibits the cAMP-synthesizing enzyme adenylate cyclase, and chelation of zinc by TPEN increases cAMP generation after stimulation with the adenylate cyclase activator forskolin. Based on our in vitro results and the in vivo observations from the literature, we suggest a model in which the intracellular-free zinc concentration limits AC activity, and the decrease of zinc after 1,25D(3) treatment promotes differentiation by relieving AC inhibition. Thus, cellular zinc homeostasis acts as an endogenous modulator of monocyte differentiation.

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Section: Discussionsupporting

confidence: 85%

Cellular zinc homeostasis is a regulator in monocyte differentiation of HL-60 cells by 1α,25-dihydroxyvitamin D3

Dubben

Hönscheid

Winkler

et al. 2010

Journal of Leukocyte Biology

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“…However, TPA did not induce a consistent increase in MT levels at any of the concentrations or times tested. We also found no evidence for TPA induction of MT in the cultured human promyelocytic leukaemia-cell line HL-60 in either the undifferentiated or neutrophil-like differentiated state (Hanke et al, 1988). These data argue against a critical role for protein kinase C activation in MT induction (Imbra & Karin, 1987) or a protective role for MT in the oxidative environment of the activated neutrophil.…”

Section: Cadmium and Zinc Determinationsmentioning

confidence: 53%

“…Samples containing more than 60 DNA were discarded or re-purified. Samples were denatured and spotted on to nitrocellulose in duplicate or quadruplicate for dot-blot analysis using one or two 32P-labelled MT-specific probes [synthetic oligonucleotides complementary to the coding region of both human MT-I and MT-IT (Hanke et al, 1988)] and a normalization probe [32P-labelled (dT)18] (Harley, 1987). Specificity of the MT probes was shown with plasmids containing MT inserts (see Fig.…”

Section: Mrna Determinationmentioning

confidence: 99%

Metallothionein mRNA and protein induction by cadmium in peripheral-blood leucocytes

Harley

Menon

Rachubinski

et al. 1989

Biochemical Journal

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To help to evaluate the role of metallothionein (MT) in peripheral-blood leucocytes, we examined MT protein and mRNA levels in these cells before and after exposure to CdCl2 in culture. Protein was assayed by 109Cd2+ binding, and RNA by dot-blot hybridization. MT was induced in both lymphocytes and adherent monocytes about 10-fold with a 12 h exposure to 10 microM-CdCl2, but absolute levels were 3-fold higher in monocytes: 57 x 10(5) (+Cd2+) versus 6 x 10(5) (-Cd2+) molecules/cell for monocytes; 18 x 10(5) (+Cd2+) versus 2 x 10(5) (-Cd2+) for lymphocytes. Polymorphonuclear cells expressed relatively little MT (0.6 x 10(5) molecules/cell), and this did not change with phorbol ester stimulation or exposure to Cd2+, arguing against a direct protective role for MT in activated neutrophils. MT mRNA levels corresponded qualitatively to expression of protein in these cells. Our data provide quantitative comparisons of leucocyte MT expression and regulation in the human population. Variation in both basal and induced MT mRNA levels reflects environmental or experimental (intra-individual) and possibly genetic (inter-individual) differences.

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“…with the MI3 universal -40 primer, the reverse primer and two internal primers, S'-GGGCAGCAGGAGCAGCAGCTC-3' (oligonucleotide b) [18] and 5'-GAGCAGGGCTGTCCC-AACATCA-3' (oligonucleotide c).…”

Section: Dna Sequencingmentioning

confidence: 99%

“…Plasmid DNA was denatured and the cloned cDNA fragments sequenced on both strands by the dideoxy method [21] using the 'Sequenase Version 2.0 Kit' (U.S.B.) with the MI3 universal -40 primer, the reverse primer and two internal primers, S'-GGGCAGCAGGAGCAGCAGCTC-3' (oligonucleotide b) [18] and 5'-GAGCAGGGCTGTCCC-AACATCA-3' (oligonucleotide c).…”

Section: Dna Sequencingmentioning

confidence: 99%

Cloning and specific polymerised‐chain‐reaction amplification of a third charge‐separable human metallothionein isoform

Soumillion

Damme

Ley

1992

European Journal of Biochemistry

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Metallothionein (MT) fractions isolated from human adult liver tissue are readily separated by anion-exchange chromatography in two isoforms, MT-1 and MT-2, which differ from each other in the nature of the amino acid residue at position 11. In fetal liver tissue, the presence of a third chargeseparable MT isoform has been previously reported. We determined its partial amino acid sequence and the sequence of a cDNA clone encoding this M T form. This confirmed the existence of another human MT isoform, hereafter named MT-0, which is characterized by the presence of a negatively charged amino acid at position 8, and by a Glu23 to Lys substitution in a strictly conserved region of the protein. Taking into account these substitutions, we are able to classify human MT isoforms into three instead of two charge-separable groups, based on the nature of three amino acid residues.The unique presence of Glu8 in MT-0 enabled us to develop an MT-0-specific amplification by the polymerase chain reaction, which revealed the presence of MT-0 mRNA in adult liver RNA samples, in spite ofthe total absence of this isoform at the protein levcl. This suggests the involvement of post-transcriptional regulatory mechanisms in the expression of this fetal MT form.Metallothioneins (MT) are ubiquitous low-molecularmass, sulphydryl-rich proteins with a high affinity for essential as well as toxic trace metals, and to which possible functions in metal metabolism and detoxification have been attributed [I). In humans, MT constitute a rather complex multiprotein family, of which the exact number of closely related isoforms is still unknown. MT fractions from human liver and kidney, as well as from different cell cultures, have been isolated and separated by anion-exchange chromatography in two isoforms, MT-1 and MT-2 (reviewed in [2]). Sequence analysis has demonstrated that MT-2 fractions consist of a single polypeptide, characterized by an Asp residue at position 11, while MT-1 fractions show microheterogeneity at several positions but contain only Glyll or Val11 residues [2]. Reverse-phase (RP) HPLC of MT fractions from human adult liver revealed at least six peaks, which where identified as one MT-2 and five distinct M r -1 forms [3].At the genomic level, Southern-blot analysis showed that these proteins are encoded by a multigene family of at least 12-15 members [4], which are distributed to five autosomes [5]. The nucleotide sequence of several human MT genes has been determined and their expression pattern characterized in

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Metallothionein RNA levels in HL‐60 cells Effect of cadmium, differentiation, and protein kinase C activation

Cited by 6 publications

References 28 publications

Cellular zinc homeostasis is a regulator in monocyte differentiation of HL-60 cells by 1α,25-dihydroxyvitamin D3

Cellular zinc homeostasis is a regulator in monocyte differentiation of HL-60 cells by 1α,25-dihydroxyvitamin D3

Metallothionein mRNA and protein induction by cadmium in peripheral-blood leucocytes

Cloning and specific polymerised‐chain‐reaction amplification of a third charge‐separable human metallothionein isoform

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