Down regulation of specific binding of [20-3H]phorbol 12,13-dibutyrate and phorbol ester-induced differentiation of human promyelocytic leukemia cells

V. Solanki, Thomas J Slaga, M. Callaham, E. Huberman

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Abstract

Binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDB) to intact human promyelocytic leukemia cells susceptible (HL-60) or resistant (R-35) to phorbol ester-induced differentiation was characterized. Specific binding of [3H]PDB to both HL-60 and R-35 cells at 37°C reached a maximum within 15-20 min. Maximal specific [3H]PDB binding to HL-60 cells was followed by a decline (down regulation) of radioactivity. This down regulation was temperature dependent, because no loss of radiolabel occurred by 1 hr at 4°C. The down regulation of bound [3H]PDB seen in HL-60 cells at 37°C was not observed with R-35 cells. Prior exposure of the HL-60 cells but not of R-35 cells to 1 μM phorbol 12-myristate 13-acetate for 90 min at 37°C caused a marked reduction in the specific binding of [3H]PDB. When [3H]PDB binding was carried out at 4°C, [3H]PDB bound to both cell types in a rapid, specific, and reversible manner. At equilibrium, HL-60 and R-35 cells were found to contain almost the same number of binding sites, which had dissociation constants of about 50 nM, indicating that the failure of R-35 cells to undergo PDB-induced differentiation was not associated with any change in the affinity or in the number of [3H]PDB binding sites. These results indicate that the down regulation specific [3H]PDB binding may be a crucial early event in the control of phorbol ester-induced terminal differentiation in HL-60 cells. Furthermore, we suggest that such down regulation may be involved in other cellular and biochemical effects of phorbol diester tumor promoters.

Original languageEnglish (US)
Pages (from-to)1722-1725
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume78
Issue number3 I
StatePublished - 1981
Externally publishedYes

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Phorbol 12,13-Dibutyrate
Phorbol Esters
HL-60 Cells
Leukemia
Down-Regulation
Binding Sites
Carcinogens
Radioactivity
Acetates
Temperature

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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title = "Down regulation of specific binding of [20-3H]phorbol 12,13-dibutyrate and phorbol ester-induced differentiation of human promyelocytic leukemia cells",
abstract = "Binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDB) to intact human promyelocytic leukemia cells susceptible (HL-60) or resistant (R-35) to phorbol ester-induced differentiation was characterized. Specific binding of [3H]PDB to both HL-60 and R-35 cells at 37°C reached a maximum within 15-20 min. Maximal specific [3H]PDB binding to HL-60 cells was followed by a decline (down regulation) of radioactivity. This down regulation was temperature dependent, because no loss of radiolabel occurred by 1 hr at 4°C. The down regulation of bound [3H]PDB seen in HL-60 cells at 37°C was not observed with R-35 cells. Prior exposure of the HL-60 cells but not of R-35 cells to 1 μM phorbol 12-myristate 13-acetate for 90 min at 37°C caused a marked reduction in the specific binding of [3H]PDB. When [3H]PDB binding was carried out at 4°C, [3H]PDB bound to both cell types in a rapid, specific, and reversible manner. At equilibrium, HL-60 and R-35 cells were found to contain almost the same number of binding sites, which had dissociation constants of about 50 nM, indicating that the failure of R-35 cells to undergo PDB-induced differentiation was not associated with any change in the affinity or in the number of [3H]PDB binding sites. These results indicate that the down regulation specific [3H]PDB binding may be a crucial early event in the control of phorbol ester-induced terminal differentiation in HL-60 cells. Furthermore, we suggest that such down regulation may be involved in other cellular and biochemical effects of phorbol diester tumor promoters.",
author = "V. Solanki and Slaga, {Thomas J} and M. Callaham and E. Huberman",
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T1 - Down regulation of specific binding of [20-3H]phorbol 12,13-dibutyrate and phorbol ester-induced differentiation of human promyelocytic leukemia cells

AU - Solanki, V.

AU - Slaga, Thomas J

AU - Callaham, M.

AU - Huberman, E.

PY - 1981

Y1 - 1981

N2 - Binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDB) to intact human promyelocytic leukemia cells susceptible (HL-60) or resistant (R-35) to phorbol ester-induced differentiation was characterized. Specific binding of [3H]PDB to both HL-60 and R-35 cells at 37°C reached a maximum within 15-20 min. Maximal specific [3H]PDB binding to HL-60 cells was followed by a decline (down regulation) of radioactivity. This down regulation was temperature dependent, because no loss of radiolabel occurred by 1 hr at 4°C. The down regulation of bound [3H]PDB seen in HL-60 cells at 37°C was not observed with R-35 cells. Prior exposure of the HL-60 cells but not of R-35 cells to 1 μM phorbol 12-myristate 13-acetate for 90 min at 37°C caused a marked reduction in the specific binding of [3H]PDB. When [3H]PDB binding was carried out at 4°C, [3H]PDB bound to both cell types in a rapid, specific, and reversible manner. At equilibrium, HL-60 and R-35 cells were found to contain almost the same number of binding sites, which had dissociation constants of about 50 nM, indicating that the failure of R-35 cells to undergo PDB-induced differentiation was not associated with any change in the affinity or in the number of [3H]PDB binding sites. These results indicate that the down regulation specific [3H]PDB binding may be a crucial early event in the control of phorbol ester-induced terminal differentiation in HL-60 cells. Furthermore, we suggest that such down regulation may be involved in other cellular and biochemical effects of phorbol diester tumor promoters.

AB - Binding of [20-3H]phorbol 12,13-dibutyrate ([3H]PDB) to intact human promyelocytic leukemia cells susceptible (HL-60) or resistant (R-35) to phorbol ester-induced differentiation was characterized. Specific binding of [3H]PDB to both HL-60 and R-35 cells at 37°C reached a maximum within 15-20 min. Maximal specific [3H]PDB binding to HL-60 cells was followed by a decline (down regulation) of radioactivity. This down regulation was temperature dependent, because no loss of radiolabel occurred by 1 hr at 4°C. The down regulation of bound [3H]PDB seen in HL-60 cells at 37°C was not observed with R-35 cells. Prior exposure of the HL-60 cells but not of R-35 cells to 1 μM phorbol 12-myristate 13-acetate for 90 min at 37°C caused a marked reduction in the specific binding of [3H]PDB. When [3H]PDB binding was carried out at 4°C, [3H]PDB bound to both cell types in a rapid, specific, and reversible manner. At equilibrium, HL-60 and R-35 cells were found to contain almost the same number of binding sites, which had dissociation constants of about 50 nM, indicating that the failure of R-35 cells to undergo PDB-induced differentiation was not associated with any change in the affinity or in the number of [3H]PDB binding sites. These results indicate that the down regulation specific [3H]PDB binding may be a crucial early event in the control of phorbol ester-induced terminal differentiation in HL-60 cells. Furthermore, we suggest that such down regulation may be involved in other cellular and biochemical effects of phorbol diester tumor promoters.

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