Characterization of a 3H-GnRH method for the measurement of GnRH binding sites in human breast cancer and breast cancer cell lines

S. J. Reshkin, A. Pezzetta, V. Albarani, F. Schittulli, A. Paradiso

Research output: Contribution to journalArticlepeer-review

Abstract

The peptide hormone, GnRH has been hypothesized to play a direct antitumoral role in certain kinds of breast cancer. This direct effect is considered to function via high affinity membrane receptors to GnRH that have been demonstrated in tumors. However, no routine assay for GnRH receptors exist limiting further research in elucidating the mechanisms of GnRH action during treatment and both the clinical usefulness of this measure and the development or refinement of GnRH treatments. Part of the problem has been that previously reported procedures require expensive, difficult (for routine clinical laboratories) iodination of specific GnRH analogs. The use of iodinated substrates present the additional problems of radioactive waste disposal and safety of laboratory personal. Further, the use of different analogs complicates the comparison of data between laboratories. We present an assay that utilizes tritiated natural GnRH in order to assay the native receptor binding and reduce both safety problems and cost of assay/waste disposal. The use of nitrocellulose filters treated overnight with 1% BSA resolved the problems of high blank values. Kinetic data demonstrate that this assay is sensitive, accurate and give results comparable with other methods. We also show that the membranes derived from the tumor and the cell cultures may be frozen at -80°C for up to 90 days without affecting the results of the assay.

Original languageEnglish
Pages (from-to)371-375
Number of pages5
JournalInternational Journal of Oncology
Volume7
Issue number2
Publication statusPublished - 1995

Keywords

  • Breast cancer
  • GnRH receptors
  • MCF-7
  • MDA-MB-435
  • Preservation

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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