Expression of luteinizing hormone-releasing hormone mRNA in the human prostatic cancer cell line LNCaP.
Recent evidence suggests that LHRH or a LHRH-like peptide might be produced by human prostatic tumor cells. To test this hypothesis, we have studied whether a mRNA for LHRH is expressed in the human prostatic cancer cell line LNCaP, by means of the RT-PCR (reverse transcription-polymerase chain reaction) technique. For these experiments, mRNA was extracted from LNCaP cells, from rat hypothalami and from rat pituitaries, reverse transcribed to cDNA and amplified via the PCR utilizing a pair of oligonucleotide primers complementary to the LHRH cDNA. Following gel electrophoresis, a band of the expected size of 228 base pairs was found in LNCaP cells as well as in the rat hypothalamus, but not in the rat anterior pituitary. This 228 base pair band from LNCaP cells and from the rat hypothalamus specifically hybridized to a 32P-labeled LHRH oligonucleotide probe. The cDNA band obtained from LNCaP cells was subcloned into a plasmid vector, and the analysis of its sequence showed a complete match with the authentic human placental LHRH cDNA. These observations clearly demonstrate that a mRNA for LHRH is expressed in human prostatic cancer cells, and suggest that LHRH or a LHRH-like peptide may be produced by these cells. To test the hypothesis whether this material might act as a local growth regulating factor on tumor cell proliferation, LNCaP cells, grown in a steroid-free medium, were treated daily with a potent LHRH antagonist. After 9, 12 and 15 days, the treatment resulted in a significant increase of tumor cell proliferation. These data clearly suggest that the LHRH mRNA expressed in LNCaP cells is possibly translated into LHRH or a LHRH-like peptide which probably functions as a local growth inhibitory factor on prostatic tumor cell proliferation, by acting on LHRH receptors.
Limonta
P
P
Department of Endocrinology, University of Milano, Italy.
Dondi
D
D
Moretti
R M
RM
Fermo
D
D
Garattini
E
E
Motta
M
M
D000818:Animals; D002277:Carcinoma / Q000378:metabolism* / Q000473:pathology; D002455:Cell Division; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D007987:Gonadotropin-Releasing Hormone