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Characterization of a Temperature-Sensitive β- Endorphin-Secreting Transformed Endometrial Cell Line^*

W. I. LI, C. L. CHEN and J. Y. CHOU

Department of Large Animal Clinical Sciences, University of Florida College of Veterinary Medicine Gainesville, Florida 32610
the Human Genetics Branch, National Institute of Child Health and Human Development, National Institutes of Health Bethesda, Maryland 20892

Address all correspondence and requests for reprints to: Dr. C. L. Chen, Department of Large Animal Clinical Sciences, J-136 JHMHC, University of Florida College of Veterinary Medicine, Gainesville, Florida 32610.

Abstract

An endometrial cell line (HRE-H9) was established and characterized to study the mechanism by which gene expression of POMC-derived peptides is controlled in uterine tissues. The HRE-H9 cell line was isolated by transforming primary rabbit endometrial cell cultures, derived from hCGtreated pseudopregnant rabbits, with a temperature-sensitive A209 mutant (tsA209) simian virus 40 at a permissive temperature (33 C). The resulting cells exhibited temperature sensitivity in growth and synthesis of immunoreactive β-endorphin (irjSEND). The ir-j8-END present in the cell extracts and culture media was assayed by a specific βEND RIA. Sephadex G-50 gel filtration chromatography of the transformed cell extracts showed three peaks of jSEND immunoreactivity. The first peak eluted at the void volume, the second peak coeluted with the β-lipotropin standard, and the third peak coincided with the porcine jSEND standard. ir-j8-END was also detectable in endometrial culture media, suggesting that the transformed endometrial cells secreted POMC-derived peptides. Our data indicate that the tsA209 mutant virus-transformed endometrial cell line provides a suitable model for study of the synthesis and regulation of POMC-derived peptides in extrapituitary tissues.

Footnotes

^* Published as Florida Agricultural Experimental Station Journal Series no. 9888

Current address: Department of Physiology and Pharmacology, University of Georgia College of Veterinary Medicine, Athens, Georgia 30602.

Received July 25, 1989.

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