title: Silencing of the GnRH type 1 receptor blocks the antiproliferative effect of the GnRH agonist, leuprolide, on the androgen-independent prostate cancer cell line DU145.
abstract: This study investigated the mechanism of action of a gonadotropin-releasing hormone (GnRH) agonist, leuprolide, on proliferation of the hormone-refractory prostate cancer cell line DU145, transfected with short hairpin RNA (shRNA), to reduce expression of the GNRHR1 gene (which encodes the GnRH type 1 receptor). DU145 cell proliferation in the presence of leuprolide (10(-9) and 10(-7) M) or control medium was measured before and after GnRHR1 knockdown. Reverse transcription-polymerase chain reaction and Western blot analysis were used to measure the degree of GNRHR1 silencing. DU145 cells treated with leuprolide (10(-9) and 10(-7) M) showed significant growth inhibition compared with control-treated DU145 cells. Transfection with GNRHR1 -shRNA significantly decreased GNRHR1 expression at 48 h. DU145 cells transfected with silencing GNRHR1 -shRNA showed normal growth patterns; however, there was no significant inhibition of proliferation of DU145 cells transfected with GNRHR1 -shRNA compared with cells transfected with control-shRNA in response to leuprolide. These data demonstrated that the antiproliferative effect of leuprolide was mediated by the GnRHR1.
authors.affiliation: Department of Urology, Korea University School of Medicine, Seoul, Republic of Korea.
authors.forename: Y H
authors.initials: YH
authors.lastname: Ko
authors.forename: Y R
authors.initials: YR
authors.lastname: Ha
authors.forename: J W
authors.initials: JW
authors.lastname: Kim
authors.forename: S G
authors.initials: SG
authors.lastname: Kang
authors.forename: H A
authors.initials: HA
authors.lastname: Jang
authors.forename: S H
authors.initials: SH
authors.lastname: Kang
authors.forename: H S
authors.initials: HS
authors.lastname: Park
authors.forename: J
authors.initials: J
authors.lastname: Cheon
mesh_terms: D000728:Androgens / Q000502:physiology; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology*; D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D003470:Culture Media; D017931:DNA Primers; D020868:Gene Silencing; D007987:Gonadotropin-Releasing Hormone / Q000819:agonists*; D006801:Humans; D016729:Leuprolide / Q000494:pharmacology*; D008297:Male; D011471:Prostatic Neoplasms / Q000473:pathology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000728:Androgens; D018931:Antineoplastic Agents, Hormonal; D003470:Culture Media; D017931:DNA Primers; D007987:Gonadotropin-Releasing Hormone; D016729:Leuprolide