Antiproliferative effects of anastrozole, methotrexate, and 5-fluorouracil on endometriosis in vitro and in vivo  Charlotte Ngô, M.D., Carole Nicco, Ph.D.,

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Antiproliferative effects of anastrozole, methotrexate, and 5-fluorouracil on endometriosis in vitro and in vivo  Charlotte Ngô, M.D., Carole Nicco, Ph.D., Mahaut Leconte, M.D., Christiane Chéreau, Pharm.D., Bernard Weill, M.D., Ph.D., Frédéric Batteux, M.D., Ph.D., Charles Chapron, M.D.  Fertility and Sterility  Volume 94, Issue 5, Pages 1632-1638.e1 (October 2010) DOI: 10.1016/j.fertnstert.2009.09.031 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Basal proliferation of cells during 48 hours, without any treatment was determined by thymidine incorporation. (A) cells from patients with ovarian endometrioma versus control patients. (B) Cells from patients with deep infiltrating endometriosis versus control patients. ∗P<.05, ∗∗P<.01, ∗∗∗P<.001. Fertility and Sterility 2010 94, 1632-1638.e1DOI: (10.1016/j.fertnstert.2009.09.031) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Proliferation of cells treated during 48 hours with (A) progesterone, (B) methotrexate, and (C) anastrozole. Results are expressed as the ratio of cpm obtained from treated cells versus cpm from untreated cells. Red dotted line shows the level of untreated cells. [PG]1 = 200 μM, [PG]2 = 100 μM, [PG]3 = 25μM, [MTX]1 = 1 mM, [MTX]2 = 100 μM, [MTX]3 = 10 μM, [AN]1 = 100 μM, [AN]2 = 10 μM, [AN]3 = 1 μM. ∗P<.05, ∗∗P<.01, ∗∗∗P<.001. Fertility and Sterility 2010 94, 1632-1638.e1DOI: (10.1016/j.fertnstert.2009.09.031) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 3 (A,B) Proliferation of cells treated during 48 hours with 5-fluorouracil. (A) Cells from patients with ovarian endometriomas versus control patients. (B) Cells from patients with deep infiltrating endometriosis (DIE) versus control patients. Results are expressed as the ratio of cpm obtained from treated cells versus cpm from untreated cells. Red dotted line shows the level of untreated cells. [5-FU]1 = 10 μM, [5-FU]2 = 1 μM, [5-FU]3 = 100 nM. ∗P<.05, ∗∗P<.01, ∗∗∗P<.001. (C–G) Mouse model of endometriosis. Macroscopic implants and histology after extraction with pathology score of mice treated with PBS (C,D) or 5-fluorouracil (E,F). Original magnification ×100. (G) Pathology score. Fertility and Sterility 2010 94, 1632-1638.e1DOI: (10.1016/j.fertnstert.2009.09.031) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Purification of stromal and epithelial cells. (A) Epithelial cells and (B) stromal cells in contrast phase microscopy. (C,E) Epithelial cells and (D,F) stromal cells stained with FITC anticytokeratine antibodies (green) and Cy3 antivimentin antibodies (red). Epithelial cells show positive staining for cytokeratine and negative staining for vimentin. Stromal cells show negative staining for cytokeratine and positive staining for vimentin. Original magnification ×100. Fertility and Sterility 2010 94, 1632-1638.e1DOI: (10.1016/j.fertnstert.2009.09.031) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions