Correlation among insulin binding, degradation, and biological activity in human breast cancer cells in long term tissue culture

C. K. Osborne, Marc E Lippman, M. E. Lippman, C. R. Kahn

Research output: Contribution to journalArticle

122 Citations (Scopus)

Abstract

Insulin interaction with four human breast cancer cell lines in tissue culture was studied with respect to specific binding to receptors, degradation, and biological responsiveness. All four lines bound and degraded 125I-labeled insulin. Binding and degradation were time, temperature, and pH dependent. Unlabeled insulin competed with 125I-labeled insulin for binding to the breast cells with half-maximal inhibition of binding being observed with less than 0.6 ng/ml for three of the four lines. Other peptides competed for insulin binding in proportion to their biological potency. Scatchard analysis of the insulin binding data revealed curvilinear plots consistent with negative cooperativity, and this was confirmed by kinetic studies of dissociation. Quantitative analysis of insulin degradation revealed a similar K(m) for all four cell lines (1.0 to 2.2 x 10-7 M), whereas maximal velocities varied over a 7-fold range. Bacitracin, a polypeptide antibiotic, inhibited insulin degradation by all four lines. In one cell line, typical competitive binding data and Scatchard plots were obtained only after inhibition of degradation with bacitracin. Insulin stimulated precursor incorporation into macromolecules and fatty acids in only two of the four cell lines; with these, significant stimulation was seen with concentrations as low as 0.6 ng/ml. No correlation was found between the amount of specific binding receptor concentration, or receptor affinity and the ability of insulin to stimulate the cells, suggesting a defect distal to the hormone-receptor interaction in the two unrespnsive lines. The two responsive cell lines showed the most insulin degradation. These human cell lines should provide a useful tool for further study of the complex mechanisms of insulin action and for the study of factors that regulate growth of human breast cancer.

Original languageEnglish
Pages (from-to)94-102
Number of pages9
JournalCancer Research
Volume38
Issue number1
StatePublished - Jan 1 1978
Externally publishedYes

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Human Activities
Insulin
Breast Neoplasms
Cell Line
Bacitracin
insulin-binding peptide
Competitive Binding
Intercellular Signaling Peptides and Proteins
Breast
Fatty Acids
Hormones
Anti-Bacterial Agents
Peptides
Temperature

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Correlation among insulin binding, degradation, and biological activity in human breast cancer cells in long term tissue culture. / Osborne, C. K.; Lippman, Marc E; Lippman, M. E.; Kahn, C. R.

In: Cancer Research, Vol. 38, No. 1, 01.01.1978, p. 94-102.

Research output: Contribution to journalArticle

Osborne, C. K. ; Lippman, Marc E ; Lippman, M. E. ; Kahn, C. R. / Correlation among insulin binding, degradation, and biological activity in human breast cancer cells in long term tissue culture. In: Cancer Research. 1978 ; Vol. 38, No. 1. pp. 94-102.
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