Cytotoxic activity of gonadotropin-releasing hormone (GnRH)-pokeweed antiviral protein conjugates in cell lines expressing GnRH receptors.
Pokeweed antiviral protein (PAP), a 29-kDa ribosome-inactivating protein isolated from the leaves of Phytolacca americana, has potent cytotoxic activity once it enters the cytoplasm of a cell. It is incapable of entering cells by itself. Therefore, our objective was to determine whether a GnRH analog could be used to deliver PAP specifically to cells expressing GnRH receptors. D-Lys(6)-GnRH-Pro(9)-ethylamide was conjugated to PAP (GnRH-PAP). Chinese hamster ovary cells stably transfected with cDNA for the murine GnRH receptor and a mouse gonadotroph tumor cell line that expresses endogenous GnRH receptors (alphaT3-1 cells) were used to evaluate the cytotoxic effects of GnRH-PAP. We also examined cytotoxicity of GnRH-PAP using human endometrial, breast, and prostate cancer cell lines. Treatment of GnRH receptor-positive cells with GnRH-PAP resulted in dose-dependent cytotoxicity. Cytotoxicity of GnRH-PAP was dependent on number of GnRH receptors (r(2) = 0.871, P < 0.05) and duration of exposure of GnRH-PAP to the cells. In contrast, GnRH-PAP was not cytotoxic to Chinese hamster ovary cells not harboring GnRH receptors. Moreover, the cytotoxic activity of GnRH-PAP could be inhibited by addition of excess GnRH analog. Neither PAP nor GnRH analog alone was cytotoxic. These results suggest that GnRH analogs can be used to specifically deliver toxin molecules to cells that express GnRH receptors. Thus, a new class of biomedicines that act as hormonotoxins against cells expressing GnRH receptors provides a novel approach for inhibiting reproduction and treating cancers that are dependent on reproductive hormones.
Yang
Wei-Hsiung
WH
Animal Reproduction and Biotechnology Laboratory, Department of Physiology, Colorado State University, Fort Collins, Colorado 80523, USA.
Wieczorck
Maciej
M
Allen
Matt C
MC
Nett
Terry M
TM
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D016466:CHO Cells; D002417:Cattle; D006224:Cricetinae; D015870:Gene Expression; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism / Q000633:toxicity*; D006801:Humans; D066298:In Vitro Techniques; D009699:N-Glycosyl Hydrolases / Q000378:metabolism / Q000633:toxicity*; D010940:Plant Proteins / Q000378:metabolism / Q000633:toxicity*; D014176:Protein Biosynthesis / Q000187:drug effects; D011966:Receptors, LHRH / Q000235:genetics* / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects
D000970:Antineoplastic Agents; D010940:Plant Proteins; D011966:Receptors, LHRH; D054789:Ribosome Inactivating Proteins, Type 1; D007987:Gonadotropin-Releasing Hormone; D009699:N-Glycosyl Hydrolases; C024275:pokeweed antiviral protein