Gonadotropin-releasing hormone (GnRH) agonist leuprolide acetate and GnRH antagonist cetrorelix acetate directly inhibit leiomyoma extracellular matrix production  Joy Lynne Britten, M.D., Minnie Malik, Ph.D., Gary Levy, M.D., Mirian Mendoza, B.S., William H. Catherino, M.D., Ph.D.  Fertility and Sterility  Volume 98, Issue 5, Pages 1299-1307 (November 2012) DOI: 10.1016/j.fertnstert.2012.07.1123 Copyright © 2012 Terms and Conditions

Figure 1 (A–C) When compared with untreated leiomyoma cells, gene expression of GnRHR1 is down-regulated when leiomyoma cells are exposed to cetrorelix at 6 and 24 hours of treatment. Leiomyoma cells treated with leuprolide demonstrate a greater than fivefold increase in expression at 6 and 24 hours. At 120 hours, the level of expression of GnRHR1 approaches that of untreated leiomyoma cells treated with leuprolide. (D–F) Analysis of GnRHR1 protein production shows no change with cetrorelix treatment at 6 or 24 hours. Cetrorelix treatment at 120 hours results in a greater than twofold reduction in protein production. GnRHR1 protein is not reduced at 120 hours at a concentration of 10−5 M with leuprolide (F). *P<.05; **P<.01. Fertility and Sterility 2012 98, 1299-1307DOI: (10.1016/j.fertnstert.2012.07.1123) Copyright © 2012 Terms and Conditions

Figure 2 (A–C) When compared with untreated leiomyoma cells, leiomyoma cells treated with cetrorelix result in a decreased expression of COL1A1 at higher concentrations. Exposure of leiomyoma cells to leuprolide results in a greater than fivefold increase in the expression of collagen 1A1 at each time point observed. (D–F) Western blot analysis demonstrates no statistically significant change in collagen 1A1 protein production at 6 hours of treatment with cetrorelix (D). At 24 hours, neither drug demonstrates a significant change in collagen 1A1 protein production (E). By 120 hours, cells treated with cetrorelix show a reduction in collagen 1A1 production by greater than twofold. Cells treated with leuprolide at 10−5 M show a slightly increased collagen 1A1 production when compared with untreated leiomyoma cells (F). *P<.05; **P<.01. Fertility and Sterility 2012 98, 1299-1307DOI: (10.1016/j.fertnstert.2012.07.1123) Copyright © 2012 Terms and Conditions

Figure 3 (A–C) When compared with untreated leiomyoma cells, 6-hour exposure of leiomyoma cells to cetrorelix results in a greater than twofold decrease in the expression of fibronectin. At 24 hours there is an increase in the expression of fibronectin at low concentrations of the same drug (B). Leuprolide treatment demonstrates a concentration-dependent increase in the expression of fibronectin at 6 hours that remains apparent at both 24 and 120 hours (A–C). Treatment with cetrorelix shows a greater than twofold increase in fibronectin protein production at 6 and 24 hours (D, E). Leuprolide treatment results in no significant change in fibronectin protein production at either 24 or 120 hours (D, E). *P<.05; **P<.01. Fertility and Sterility 2012 98, 1299-1307DOI: (10.1016/j.fertnstert.2012.07.1123) Copyright © 2012 Terms and Conditions

Figure 4 (A–C) When compared with untreated leiomyoma cells, cells treated with cetrorelix result in a marked decrease in the gene expression of versican variant V0 at both 6 and 24 hours, whereas leuprolide treatment increases versican V0 expression at 6 and 24 hours by greater than 10-fold. (D–F) At 6 hours, versican variant V0 protein production decreases slightly at lower concentrations when exposed to cetrorelix. At 24 hours, versican variant V0 production seems unchanged with either drug treatment (E). However, when leiomyoma cells are exposed to cetrorelix at 120 hours, versican variant V0 protein production is initially increased, but begins to approach that of untreated leiomyoma cells at 10−7 M. Leuprolide-treated leiomyoma cells demonstrate no change in protein production at 120 hours of treatment. *P<.05; **P<.01. Fertility and Sterility 2012 98, 1299-1307DOI: (10.1016/j.fertnstert.2012.07.1123) Copyright © 2012 Terms and Conditions