Epidermal growth factor receptors in clonal lines of a rat osteogenic sarcoma and in osteoblast-rich rat bone cells

K. W. Ng, N. C. Partridge, M. Niall, T. J. Martin

Research output: Contribution to journalArticlepeer-review

Abstract

Studies were carried out to identify and characterize the receptors for epidermal growth factor (EGF) in osteoblast-rich newborn rat calvarial cells and in 4 clonal lines derived from a transplantable rat osteogenic sarcoma with a well-characterized osteoblast-like phenotype. The cells were grown in monolayer culture in replicate wells; 40,000-50,000 cpm125I-labeled mouse EGF with a specific activity of 100-120 μCi/μg was added to each well. Binding studies were carried out at 37°C. Binding of125I-labeled EGF was specific, saturable, reversible, and pH dependent. Maximum binding occurred 2 h after addition of the tracer. Thereafter, cell-bound radioactivity decreased to reach a plateau of 15-20% of maximum binding at 24 h. This observation is consistent with internalization and processing of the receptor-hormone complex as has been shown with other EGF target cells. Scatchard analyses revealed a single class of high-affinity binding sites in the normal and malignant osteoblast-like cells. Dissociation constants (KD) in the clonal lines ranged from 2.3 × 10-10M to 4.7 × 10-10M with receptor number per cell ranging from 25,000 to 33,000. The calvarial cells had a KD of 2.0 × 10-10M with 14,000 receptors per cell. In both the normal and malignant cell strains, EGF was found to increase incorporation of3H-labeled thymidine into acid-precipitable macromolecules. EGF has been shown to stimulate bone resorption; however, studies in organ cultures have not identified the target cell for EGF. The present results point to an interaction of EGF with osteoblasts.

Original languageEnglish (US)
Pages (from-to)298-303
Number of pages6
JournalCalcified Tissue International
Volume35
Issue number1
DOIs
StatePublished - Dec 1983

Keywords

  • Epidermal growth factor
  • Osteoblasts
  • Receptors

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine
  • Endocrinology

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