Characterization of the association of radiolabeled bleomycin A2 with HeLa cells

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

The association of [3H]bleomycin A2 and Cu(II):[3H]bleomycin A2 with HeLa cells has been characterized. Under the conditions of our experiments, approximately 0.1% of the total drug in the medium associates with HeLa cells. Both forms of the drug bind to HeLa cells in a specific and saturable manner, with a K(m) of 20 μM and a V(max) of 2.5 pmol/min/106 cells. Scatchard analysis of the specific binding data demonstrates a single set of high-affinity binding sites. Cytotoxic activities of both forms of the drug are similar, with a 50% lethal dose of 0.5 μM at 48 hr. The specific binding in HeLa cells of either the labeled metal-free drug or its copper complex is reversible by a 100-fold excess of either unlabeled drug. Interaction of the drug with cells is temperature sensitive but is unaffected by metabolic poisons, suggesting that this process is not energy dependent. Isolation of DNA from HeLa cells incubated with the drug indicates that 1 mol of either [3H]bleomycin A2 or Cu(II):[3H]bleomycin A2 binds per 108 nucleotides. Further studies with the radiolabeled drug are required to define precisely the mechanism involved in bleomycin uptake and compartmentalization within the cell.

Original languageEnglish (US)
Pages (from-to)1541-1546
Number of pages6
JournalCancer Research
Volume44
Issue number4
StatePublished - 1984

Fingerprint

Bleomycin
HeLa Cells
Pharmaceutical Preparations
Poisons
Lethal Dose 50
Drug Interactions
Copper
Nucleotides
Metals
Binding Sites
Temperature
DNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Characterization of the association of radiolabeled bleomycin A2 with HeLa cells. / Roy, S. N.; Band Horwitz, Susan.

In: Cancer Research, Vol. 44, No. 4, 1984, p. 1541-1546.

Research output: Contribution to journalArticle

@article{c6103b59a2ac4cc4ac15c6fb352c8317,
title = "Characterization of the association of radiolabeled bleomycin A2 with HeLa cells",
abstract = "The association of [3H]bleomycin A2 and Cu(II):[3H]bleomycin A2 with HeLa cells has been characterized. Under the conditions of our experiments, approximately 0.1{\%} of the total drug in the medium associates with HeLa cells. Both forms of the drug bind to HeLa cells in a specific and saturable manner, with a K(m) of 20 μM and a V(max) of 2.5 pmol/min/106 cells. Scatchard analysis of the specific binding data demonstrates a single set of high-affinity binding sites. Cytotoxic activities of both forms of the drug are similar, with a 50{\%} lethal dose of 0.5 μM at 48 hr. The specific binding in HeLa cells of either the labeled metal-free drug or its copper complex is reversible by a 100-fold excess of either unlabeled drug. Interaction of the drug with cells is temperature sensitive but is unaffected by metabolic poisons, suggesting that this process is not energy dependent. Isolation of DNA from HeLa cells incubated with the drug indicates that 1 mol of either [3H]bleomycin A2 or Cu(II):[3H]bleomycin A2 binds per 108 nucleotides. Further studies with the radiolabeled drug are required to define precisely the mechanism involved in bleomycin uptake and compartmentalization within the cell.",
author = "Roy, {S. N.} and {Band Horwitz}, Susan",
year = "1984",
language = "English (US)",
volume = "44",
pages = "1541--1546",
journal = "Cancer Research",
issn = "0008-5472",
publisher = "American Association for Cancer Research Inc.",
number = "4",

}

TY - JOUR

T1 - Characterization of the association of radiolabeled bleomycin A2 with HeLa cells

AU - Roy, S. N.

AU - Band Horwitz, Susan

PY - 1984

Y1 - 1984

N2 - The association of [3H]bleomycin A2 and Cu(II):[3H]bleomycin A2 with HeLa cells has been characterized. Under the conditions of our experiments, approximately 0.1% of the total drug in the medium associates with HeLa cells. Both forms of the drug bind to HeLa cells in a specific and saturable manner, with a K(m) of 20 μM and a V(max) of 2.5 pmol/min/106 cells. Scatchard analysis of the specific binding data demonstrates a single set of high-affinity binding sites. Cytotoxic activities of both forms of the drug are similar, with a 50% lethal dose of 0.5 μM at 48 hr. The specific binding in HeLa cells of either the labeled metal-free drug or its copper complex is reversible by a 100-fold excess of either unlabeled drug. Interaction of the drug with cells is temperature sensitive but is unaffected by metabolic poisons, suggesting that this process is not energy dependent. Isolation of DNA from HeLa cells incubated with the drug indicates that 1 mol of either [3H]bleomycin A2 or Cu(II):[3H]bleomycin A2 binds per 108 nucleotides. Further studies with the radiolabeled drug are required to define precisely the mechanism involved in bleomycin uptake and compartmentalization within the cell.

AB - The association of [3H]bleomycin A2 and Cu(II):[3H]bleomycin A2 with HeLa cells has been characterized. Under the conditions of our experiments, approximately 0.1% of the total drug in the medium associates with HeLa cells. Both forms of the drug bind to HeLa cells in a specific and saturable manner, with a K(m) of 20 μM and a V(max) of 2.5 pmol/min/106 cells. Scatchard analysis of the specific binding data demonstrates a single set of high-affinity binding sites. Cytotoxic activities of both forms of the drug are similar, with a 50% lethal dose of 0.5 μM at 48 hr. The specific binding in HeLa cells of either the labeled metal-free drug or its copper complex is reversible by a 100-fold excess of either unlabeled drug. Interaction of the drug with cells is temperature sensitive but is unaffected by metabolic poisons, suggesting that this process is not energy dependent. Isolation of DNA from HeLa cells incubated with the drug indicates that 1 mol of either [3H]bleomycin A2 or Cu(II):[3H]bleomycin A2 binds per 108 nucleotides. Further studies with the radiolabeled drug are required to define precisely the mechanism involved in bleomycin uptake and compartmentalization within the cell.

UR - http://www.scopus.com/inward/record.url?scp=0021267030&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021267030&partnerID=8YFLogxK

M3 - Article

C2 - 6200212

AN - SCOPUS:0021267030

VL - 44

SP - 1541

EP - 1546

JO - Cancer Research

JF - Cancer Research

SN - 0008-5472

IS - 4

ER -