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In vitro evaluation of the growth inhibition and apoptosis effect of mifepristone (RU486) in human Ishikawa and HEC1A endometrial cancer cell lines

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Abstract

Purpose

To determine the growth inhibitory effects of mifepristone on endometrial cancer cell growth and evaluate its effect on apoptosis using HEC-1-A and Ishikawa human endometrial cancer cell lines.

Methods

The human endometrial cancer cell lines, HEC-1-A and Ishikawa, were cultured in vitro. MTT assays were completed in order to estimate the IC50 of mifepristone. Both cell lines were then treated with the respective IC50 values. Immunohistochemistry assays were performed to investigate the expression of estrogen receptors alpha and beta (ERα/β), progesterone receptor alpha and beta (PR α/β), cyclooxygenase-2 (COX-2), bax, p53, and bcl-2. Flow cytometry analysis was performed to study cell cycle arrest and apoptosis.

Results

The estimated IC50 of mifepristone for HEC-1-A and Ishikawa was found to be 16 and 19 μg/ml respectively. At this concentration, there was no change in either ERα/β or PR α/β in Ishikawa. However, PR β expression increased with time of treatment in HEC-1-A. Expression of p53 was increased with duration of treatment in both cell lines. Consequently a decrease in bcl-2 and an increase in COX-2 expression were seen in HEC-1-A and Ishikawa cells, respectively. Lastly, flow cytometry analysis confirmed an accumulation of cells in G0 phase after 72 h of treatment in both cell lines.

Conclusions

Mifepristone demonstrates activity in both HEC-1-A and Ishikawa cells at clinically relevant concentrations based on an oral human dose of about 200 mg/day. While its mechanism of action remains unknown, this data supports an increase in apoptosis that may be due to p53 activation rather than hormone receptor mediation. Additional studies are needed to help further identify mifepristone mechanism of action.

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Authors and Affiliations

  1. Division of Pharmacy, The University of Texas, M. D. Anderson Cancer Center, Houston, TX, USA

    Marisa A. Navo & Judith A. Smith

  2. Department of Gynecologic Oncology, Division of Surgery, The University of Texas, M. D. Anderson Cancer Center, Unit 1362, PO Box 301439, Houston, TX, 77230-1439, USA

    Judith A. Smith, Anjali Gaikwad, Thomas Burke, Jubilee Brown & Lois M. Ramondetta

  3. Department of Obstetrics and Gynecology, University of Texas Health Sciences Center at Houston Medical School, Houston, TX, USA

    Judith A. Smith

Authors
  1. Marisa A. Navo
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  2. Judith A. Smith
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  3. Anjali Gaikwad
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  4. Thomas Burke
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  5. Jubilee Brown
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  6. Lois M. Ramondetta
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Correspondence to Lois M. Ramondetta.

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Navo, M.A., Smith, J.A., Gaikwad, A. et al. In vitro evaluation of the growth inhibition and apoptosis effect of mifepristone (RU486) in human Ishikawa and HEC1A endometrial cancer cell lines. Cancer Chemother Pharmacol 62, 483–489 (2008). https://doi.org/10.1007/s00280-007-0628-z

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  • DOI: https://doi.org/10.1007/s00280-007-0628-z

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