Characterization of differentiation-inducer-resistant HL-60 cells

Robert E. Gallagher, Patricia A. Bilello, Anna C. Ferrari, Chin Sen Chang, Ray-Whay Yen, Wayne A. Nickols, Emil C. Muly

Research output: Contribution to journalArticle

Abstract

Sub-lines of the cultured human promyelocytic leukemia cell line HL-60 were individually selected for their ability to sustain exponential growth in the presence of 3 structurallyunrelated inducers of granulocytic differentiation - retinoic acid (RA), dimethylsulfoxide (DMSO), and 6-thioguanine (6TG). Selections were made by step-wise augmentation to final drug concentrations of 10-3mM RA, 169mM (1.2%) DMSO and 0.12mM (20 μg ml-1) 6TG. In addition to growth resistance, cells in each sub-line displayed variable cytodifferentiation resistance to each of the 3 selective agents, which was quantitated as the ratio of the concentration of drug required to induce differentiation in 50% of the cells in each resistant sub-line versus comparably-passaged wild-type HL-60 cells. The levels of resistance/cross-resistance were as follows: RA-resistant (res) sub-line > 2700-fold to RA, 1.3-fold to DMSO and > 1.5-fold to hypoxanthine (HXN; the noncytotoxic purine base inducer analogue of 6TG); DMSO-res sub-line 2.5-fold to DMSO, 137-fold to RA and > 1.5-fold to HXN; and 6TG-res sub-line > 1.5-fold to HXN, 9-fold to RA and 1.6-fold to DMSO. These sub-lines were not cross-resistant to sodium butyrate (NaBut), a monocyte inducer, or to 12-O-tetradecanoylphorbol 13-acetate (TPA), a macrophage inducer. HL-60 sub-lines selected by exposure to a single high concentration of 5-bromo-2′-deoxyuridine (BUdR; 3.3 × 10-2mM) or oubain (Ou; 5 × 10-3mM) were not or were slightly cross-resistant to either granulocyte or monocyte inducers. Although some variations in line/sub-line phenotype were observed, this was minor compared to the quantitative variations in response to individual inducing agents. The RA-res and 6TG-res sub-lines contained numerous double minute chromosomes (indicators of amplified genes) which were either absent or present in much smaller numbers in the parental wild-type cells or in the other drug-resistant sub-lines. There was little change or a decrease in the amplification level of the known amplified oncogene c-myc in the various drug-resistant sub-lines compared to wild-type HL-60 cells. These results (a) confirm that the neutrophilic granulocytic and monocytic/macrophagic differentiation programs in HL-60 cells are mechanistically different and separable; (b) suggest that both agent-specific and common quantitative alterations contribute to the mechanism(s) for resistance to granulocyte differentiation; and (c) suggest that the latter quantitative defects could be related to amplification of genes other than c-myc.

Original languageEnglish (US)
JournalLeukemia Research
Volume9
Issue number8
DOIs
StatePublished - 1985
Externally publishedYes

Fingerprint

HL-60 Cells
Tretinoin
Thioguanine
Dimethyl Sulfoxide
myc Genes
Bromodeoxyuridine
Granulocytes
Pharmaceutical Preparations
Monocytes
Hypoxanthine
Butyric Acid
Tetradecanoylphorbol Acetate
Growth
Leukemia
Chromosomes
Macrophages
Phenotype
Cell Line
Genes

Keywords

  • double minute chromosomes
  • drug/differentiation resistance
  • gene amplification
  • Myeloid leukemic cell differentiation
  • oncogenes

ASJC Scopus subject areas

  • Cancer Research
  • Hematology
  • Oncology

Cite this

Gallagher, R. E., Bilello, P. A., Ferrari, A. C., Chang, C. S., Yen, R-W., Nickols, W. A., & Muly, E. C. (1985). Characterization of differentiation-inducer-resistant HL-60 cells. Leukemia Research, 9(8). https://doi.org/10.1016/0145-2126(85)90067-0

Characterization of differentiation-inducer-resistant HL-60 cells. / Gallagher, Robert E.; Bilello, Patricia A.; Ferrari, Anna C.; Chang, Chin Sen; Yen, Ray-Whay; Nickols, Wayne A.; Muly, Emil C.

In: Leukemia Research, Vol. 9, No. 8, 1985.

Research output: Contribution to journalArticle

Gallagher, RE, Bilello, PA, Ferrari, AC, Chang, CS, Yen, R-W, Nickols, WA & Muly, EC 1985, 'Characterization of differentiation-inducer-resistant HL-60 cells', Leukemia Research, vol. 9, no. 8. https://doi.org/10.1016/0145-2126(85)90067-0
Gallagher, Robert E. ; Bilello, Patricia A. ; Ferrari, Anna C. ; Chang, Chin Sen ; Yen, Ray-Whay ; Nickols, Wayne A. ; Muly, Emil C. / Characterization of differentiation-inducer-resistant HL-60 cells. In: Leukemia Research. 1985 ; Vol. 9, No. 8.
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AU - Gallagher, Robert E.

AU - Bilello, Patricia A.

AU - Ferrari, Anna C.

AU - Chang, Chin Sen

AU - Yen, Ray-Whay

AU - Nickols, Wayne A.

AU - Muly, Emil C.

PY - 1985

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N2 - Sub-lines of the cultured human promyelocytic leukemia cell line HL-60 were individually selected for their ability to sustain exponential growth in the presence of 3 structurallyunrelated inducers of granulocytic differentiation - retinoic acid (RA), dimethylsulfoxide (DMSO), and 6-thioguanine (6TG). Selections were made by step-wise augmentation to final drug concentrations of 10-3mM RA, 169mM (1.2%) DMSO and 0.12mM (20 μg ml-1) 6TG. In addition to growth resistance, cells in each sub-line displayed variable cytodifferentiation resistance to each of the 3 selective agents, which was quantitated as the ratio of the concentration of drug required to induce differentiation in 50% of the cells in each resistant sub-line versus comparably-passaged wild-type HL-60 cells. The levels of resistance/cross-resistance were as follows: RA-resistant (res) sub-line > 2700-fold to RA, 1.3-fold to DMSO and > 1.5-fold to hypoxanthine (HXN; the noncytotoxic purine base inducer analogue of 6TG); DMSO-res sub-line 2.5-fold to DMSO, 137-fold to RA and > 1.5-fold to HXN; and 6TG-res sub-line > 1.5-fold to HXN, 9-fold to RA and 1.6-fold to DMSO. These sub-lines were not cross-resistant to sodium butyrate (NaBut), a monocyte inducer, or to 12-O-tetradecanoylphorbol 13-acetate (TPA), a macrophage inducer. HL-60 sub-lines selected by exposure to a single high concentration of 5-bromo-2′-deoxyuridine (BUdR; 3.3 × 10-2mM) or oubain (Ou; 5 × 10-3mM) were not or were slightly cross-resistant to either granulocyte or monocyte inducers. Although some variations in line/sub-line phenotype were observed, this was minor compared to the quantitative variations in response to individual inducing agents. The RA-res and 6TG-res sub-lines contained numerous double minute chromosomes (indicators of amplified genes) which were either absent or present in much smaller numbers in the parental wild-type cells or in the other drug-resistant sub-lines. There was little change or a decrease in the amplification level of the known amplified oncogene c-myc in the various drug-resistant sub-lines compared to wild-type HL-60 cells. These results (a) confirm that the neutrophilic granulocytic and monocytic/macrophagic differentiation programs in HL-60 cells are mechanistically different and separable; (b) suggest that both agent-specific and common quantitative alterations contribute to the mechanism(s) for resistance to granulocyte differentiation; and (c) suggest that the latter quantitative defects could be related to amplification of genes other than c-myc.

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KW - Myeloid leukemic cell differentiation

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