In vitro effects of phthalate esters in human myometrial and leiomyoma cells and increased urinary level of phthalate metabolite in women with uterine.

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In vitro effects of phthalate esters in human myometrial and leiomyoma cells and increased urinary level of phthalate metabolite in women with uterine leiomyoma Jin Hee Kim, Ph.D., Sung Hoon Kim, M.D., Ph.D., Young Sang Oh, M.S., Hyo Jin Ihm, M.S., Hee Dong Chae, M.D., Ph.D., Chung-Hoon Kim, M.D., Ph.D., Byung Moon Kang, M.D., Ph.D. Fertility and Sterility Volume 107, Issue 4, Pages 1061-1069.e1 (April 2017) DOI: 10.1016/j.fertnstert.2017.01.015 Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Cellular viability of human myometrial cells (A), leiomyoma cells (B), estrogen receptor–positive breast cancer (MCF-7) cells (C), and estrogen receptor–negative breast cancer (MD-MB-468) cells (D) after exposure to DEHP. Cells were exposed to DEHP for 24–72 hours at concentrations of 10−8 M and 10−6 M. Error bars show the mean ± SEM. *P<.05 vs. control. con = control; P6 = DEHP 10−6 M; P8 = DEHP 10−8 M. Fertility and Sterility 2017 107, 1061-1069.e1DOI: (10.1016/j.fertnstert.2017.01.015) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Effects of in vitro DEHP treatment on cellular proliferation in human myometrial and leiomyoma cells. Proliferating cell nuclear antigen expression levels were measured by Western blot analyses in human myometrial cells (A) and leiomyoma cells (B). Ki-67 expression levels were measured by immunofluorescence staining in human myometrial cells (C) and leiomyoma cells (D). Western blot data are expressed as a percentage, whereby cells treated with vehicle are normalized to 100%. Error bars show the mean ± SEM. *P<.05 vs. control. con = control; P6 = DEHP 10−6 M; P8 = DEHP 10−8 M. Fertility and Sterility 2017 107, 1061-1069.e1DOI: (10.1016/j.fertnstert.2017.01.015) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Effects of in vitro DEHP treatment on Bcl-2 expression and collagen type I protein expression in human myometrial and leiomyoma cells. Bcl-2 expression levels were measured by Western blot analyses in human myometrial cells (A) and leiomyoma cells (B). Collagen type I expression levels were measured by Western blot analyses in human myometrial cells (C) and leiomyoma cells (D). Data are expressed as a percentage, whereby cells treated with vehicle are normalized to 100%. Error bars show the mean ± SEM. *P<.05 vs. control. con = control; P6 = DEHP 10−6 M; P8 = DEHP 10−8 M. Fertility and Sterility 2017 107, 1061-1069.e1DOI: (10.1016/j.fertnstert.2017.01.015) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Immunocytochemistry of human myometrial cells (A–C) and leiomyoma cells (D–F). Human myometrial cells and uterine leiomyoma cells stained for α-SMA (A, D) and vimentin (B, E). (C, F) Negative controls. Original magnification, ×200. Fertility and Sterility 2017 107, 1061-1069.e1DOI: (10.1016/j.fertnstert.2017.01.015) Copyright © 2017 American Society for Reproductive Medicine Terms and Conditions