title: Peptidomimetic GnRH antagonist AEZS-115 inhibits the growth of ovarian and endometrial cancer cells.
abstract: BACKGROUND
  AEZS-115 (Aeterna Zentaris GmbH, Frankfurt/M, Germany) is an orally active peptidomimetic antagonist of gonadotropin-releasing hormone (GnRH). In various tumors, an autocrine growth-promoting loop has been described for GnRH. The current study evaluates the antitumor activity and mechanism of action of AEZS-115 in models of ovarian and endometrial cancer.
  
  
  METHODS
  Human A2780, Acis2780, OAW-42, Ovcar-3, SKOV-3, Hec1A and Ishikawa cells were analyzed for GnRH receptor expression by reverse transcription polymerase chain reaction (RT-PCR). These cell lines were incubated with AEZS-115 at 1, 10 and 100 μM for 24 h, 48 h, and 72 h and the number of viable cells was determined. Fluorescence activated cell sorting (FACS) cell cycle analyses were performed with increasing concentrations of AEZS-115. Co-treatment experiments of cancer cells with GnRH antagonist cetrorelix and peptidomimetic GnRH antagonist AESZ-115 were carried out.
  
  
  RESULTS
  A2780, Acis2780, OAW-42, Ovcar-3, SKOV-3, Hec1A and Ishikawa cells expressed GnRH receptors as demonstrated by RT-PCR. GnRH antagonist AEZS-115 inhibited growth of all cell lines in a dose- and time-dependent manner. Half maximal inhibitory concentration (IC(50)) values at 48 h of incubation were between 7 and 17.5 μM and for 72 h between 4.5 and 12.5 μM. IC(50) values for ovarian and endometrial cancer cells were rather similar. These results were obtained by tetrazolium salt [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; MTT] assay and confirmed by additional crystal violet staining. Cell cycle FACS analysis revealed that AEZS-115 dose-dependently increased the fraction of apoptotic cells. Co-treatment experiments carried out with AEZS-115 and peptidic GnRH-antagonist cetrorelix suggest that the antitumor effect of AEZS-115 is not mediated by blockade of the GnRH receptor.
  
  
  CONCLUSIONS
  GnRH antagonist AEZS-115 exhibited substantial antitumor activity in ovarian as well as endometrial cancer cell lines. However, this antitumor effect was not mediated by the tumoral GnRH receptors. To identify the mechanism of action of this compound, further research is warranted. Its in vitro antitumor activity makes AEZS-115 a promising candidate for in vivo studies of ovarian and endometrial cancer.
authors.affiliation: Department of Gynecology and Obstetrics, Medical University of Würzburg, Josef-Schneider-Str. 4, 97080 Würzburg, Germany. joergbengel@hotmail.com
authors.forename: J B
authors.initials: JB
authors.lastname: Engel
authors.forename: J C
authors.initials: JC
authors.lastname: Hahne
authors.forename: S F M
authors.initials: SF
authors.lastname: Häusler
authors.forename: S
authors.initials: S
authors.lastname: Meyer
authors.forename: S E
authors.initials: SE
authors.lastname: Segerer
authors.forename: J
authors.initials: J
authors.lastname: Diessner
authors.forename: J
authors.initials: J
authors.lastname: Dietl
authors.forename: A
authors.initials: A
authors.lastname: Honig
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D016889:Endometrial Neoplasms / Q000188:drug therapy* / Q000473:pathology; D005260:Female; D007987:Gonadotropin-Releasing Hormone / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D006801:Humans; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000473:pathology; D057786:Peptidomimetics / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D011966:Receptors, LHRH / Q000235:genetics
chemical_list: D000590:Amino Acid Chloromethyl Ketones; C484844:GNRHR protein, human; D057786:Peptidomimetics; D012333:RNA, Messenger; D011966:Receptors, LHRH; C484771:benzyloxycarbonyl-valyl-aspartic acid fluoromethyl ketone; D007987:Gonadotropin-Releasing Hormone; C062876:cetrorelix