Steroid Sulfatase Inhibitor Reduces Proliferation of Ishikawa Endometrial Cancer Cells in Co-Culture Systems

DOI: 10.4236/ojemd.2016.69025   PDF   HTML   XML   1,314 Downloads   1,806 Views  


Objectives: Estrogens significantly contribute toward the growth and development of endometrial cancers. Two principal pathways have been implicated in the final steps of estrogen synthesis: the steroid sulfatase (STS) and aromatase pathways. In this study, we aimed to evaluate the possible effects of tumor-stromal interactions on local estrogen biosynthesis in endometrial cancer. We also assessed the biological effects of inhibitors of steroid sulfatase and aromatase in the co-culture system compared with usual monocultures. Methods/Materials: We isolated stromal cells from endometrial cancer patients to examine local biosynthesis of estrogens and tumor-stromal interactions. Next we examined the effects of steroid sulfatase inhibitor and aromatase inhibitor in monoculture of endometrial cancer cell line (Ishikawa) and in a co-culture system involving an Ishikawa cells and stromal cells. Results: Estrogen receptor and steroid sulfatase mRNA levels in cancer cells were significantly higher in the co-cultures compared with the monocultures of endometrial cancer cells. Estradiol and androstenediol concentrations were also significantly higher in the co-cultured cells. Proliferation of the cancer cells was significantly increased through the steroid sulfatase pathway, which metabolizes androgens, estrone sulfate, and estradiol sulfate as its substrates. However, its proliferation was significantly decreased by the treatment of steroid sulfatase or aromatase inhibitors. The significant growth inhibition by the steroid sulfatase and aromatase inhibitors were also observed in the co-culture system. Conclusions: We evaluated the effects of STS inhibitor and aromatase inhibitors on the proliferation of estrogen-dependent endometrial cancer cells. Considering that intratumoral estrogen metabolism plays an important role, our co-culture systems provide an environment similar to that of the tumor in living patients in terms of metabolism and synthesis of intratumoral estrogens. The results of this study may aid in achieving improved clinical responses from patients treated with STS inhibitors.

Share and Cite:

Nishimoto, M. , Toyoshima, M. , Shiga, N. , Utsunomiya, H. , Suzuki, F. , Nagase, S. , Nishigori, H. , Suzuki, T. , Sasano, H. , Ito, K. and Yaegashi, N. (2016) Steroid Sulfatase Inhibitor Reduces Proliferation of Ishikawa Endometrial Cancer Cells in Co-Culture Systems. Open Journal of Endocrine and Metabolic Diseases, 6, 193-204. doi: 10.4236/ojemd.2016.69025.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Siegel, R., Naishadham, D. and Jemal, A. (2012) Cancer Statistics, 2012. CA: A Cancer Jour- nal for Clinicians, 62, 10-29.
[2] Berstein, L.M., Tchernobrovkina, A.E., Gamajunova, V.B., Kovalevskij, A.J., Vasilyev, D.A., Chepik, O.F., et al. (2003) Tumor Estrogen Content and Clinico-Morphological and Endocrine Features of Endometrial Cancer. Journal of Cancer Research and Clinical Oncology, 129, 245-249.
[3] Ito, K., Utsunomiya, H., Niikura, H., Yaegashi, N. and Sasano, H. (2011) Inhibition of Estrogen Actions in Human Gynecological Malignancies: New Aspects of Endocrine Therapy for Endometrial Cancer and Ovarian Cancer. Molecular and Cellular Endocrinology, 340, 161-167.
[4] Bulun, S.E., Lin, Z., Imir, G., Amin, S., Demura, M., Yilmaz, B., et al. (2005) Regulation of Aromatase Expression in Estrogen-Responsive Breast and Uterine Disease: From Bench to Treatment. Pharmacological Reviews, 57, 359-383.
[5] Adessi, G.L., Prost, O., Agnani, G., Petitjean, A. and Burnod, J. (1984) Estrone Sulfatase Activity in Normal and Abnormal Endometrium. Archives of Gynecology, 236, 13-18.
[6] Bonney, R.C., Scanlon, M.J., Jones, D.L., Reed, M.J., Anderson, M.C. and James, V.H. (1986) The Relationship between Oestradiol Metabolism and Adrenal Steroids in the Endometrium of Postmenopausal Women with and without Endometrial Cancer. European Journal of Cancer and Clinical Oncology, 22, 953-961.
[7] Tseng, L., Mazella, J., Mann, W.J. and Chumas, J. (1982) Estrogen Synthesis in Normal and Malignant Human Endometrium. The Journal of Clinical Endocrinology & Metabolism, 55, 1029-1031.
[8] Cornel, K.M.C., Kruitwagen, R.F.P.M., Delvoux, B., Visconti, L., Van de Vijver, K.K., Day, J.M., et al. (2012) Overexpression of 17β-Hydroxysteroid Dehydrogenase Type 1 Increases the Exposure of Endometrial Cancer to 17β-Estradiol. The Journal of Clinical Endocrinology & Metabolism, 97, E591-E601.
[9] Hevir, N., Sinkovec, J. and Ri?ner, T.L. (2011) Disturbed Expression of Phase I and Phase II Estrogen-Metabolizing Enzymes in Endometrial Cancer: Lower Levels of CYP1B1 and Increased Expression of S-COMT. Molecular and Cellular Endocrinology, 331, 158-167.
[10] Jongen, V.H.W.M., Bri?t, J.M., de Jong, R.A., Joppe, E., ten Hoor, K.A., Boezen, H.M., et al. (2009) Aromatase, Cyclooxygenase 2, HER-2/neu, and p53 as Prognostic Factors in Endometrioid Endometrial Cancer. International Journal of Gynecological Cancer, 19, 670-676.
[11] Pathirage, N., Di Nezza, L.A., Salmonsen, L.A., Jobling, T., Simpson, E.R. and Clyne, C.D. (2006) Expression of Aromatase, Estrogen Receptors, and Their Coactivators in Patients with Endometrial Cancer. Fertility Sterility, 86, 469-472.
[12] Sinreih, M., Hevir, N. and Ri?ner, T.L. (2013) Altered Expression of Genes Involved in Progesterone Biosynthesis, Metabolism and Action in Endometrial Cancer. Chemico-Bio- logical Interactions, 202, 210-217.
[13] Smuc, T. and Rizner, T.L. (2009) Aberrant Pre-Receptor Regulation of Estrogen and Progesterone Action in Endometrial Cancer. Molecular and Cellular Endocrinology, 301, 74-82.
[14] Moeller, G. and Adamski, J. (2009) Integrated View on 17beta-Hydroxysteroid Dehydrogenases. Molecular and Cellular Endocrinology, 301, 7-19.
[15] Suzuki, T., Nakata, T., Miki, Y., Kaneko, C., Moriya, T., Ishida, T., et al. (2003) Estrogen Sulfotransferase and Steroid Sulfatase in Human Breast Carcinoma. Cancer Research, 63, 2762-2770.
[16] Utsumi, T., Yoshimura, N., Takeuchi, S., Maruta, M., Maeda, K. and Harada, N. (2000) Elevated Steroid Sulfatase Expression in Breast Cancers. Journal of Steroid Biochemistry and Molecular Biology, 73, 141-145.
[17] Suzuki, T., Miki, Y., Nakamura, Y., Ito, K. and Sasano, H. (2011) Steroid Sulfatase and Estrogen Sulfotransferase in Human Carcinomas. Molecular and Cellular Endocrinology, 340, 148-153.
[18] Utsunomiya, H., Ito, K., Suzuki, T., Kitamura, T., Kaneko, C., Nakata, T., et al. (2004) Steroid Sulfatase and Estrogen Sulfotransferase in Human Endometrial Carcinoma. Clinical Cancer Research, 10, 5850-5856.
[19] Yamaguchi, Y., Takei, H., Suemasu, K., Kobayashi, Y., Kurosumi, M., Harada, N., et al. (2005) Tumor-Stromal Interaction through the Estrogen-Signaling Pathway in Human Breast Cancer. Cancer Research, 65, 4653-4562.
[20] Takahashi-Shiga, N., Utsunomiya, H., Miki, Y., Nagase, S., Kobayashi, R., Matsumoto, M., et al. (2009) Local Biosynthesis of Estrogen in Human Endometrial Carcinoma through tumor-Stromal Cell Interactions. Clinical Cancer Research, 15, 6028-6034.
[21] Miki, Y., Suzuki, T., Tazawa, C., Yamaguchi, Y., Kitada, K., Honma, S., et al. (2007) Aromatase Localization in Human Breast Cancer Tissues: Possible Interactions between Intratumoral Stromal and Parenchymal Cells. Cancer Research, 67, 3945-3954.
[22] Matsumoto, M., Yamaguchi, Y., Seino, Y., Hatakeyama, A., Takei, H., Niikura, H., et al. (2008) Estrogen Signaling Ability in Human Endometrial Cancer through the Cancer-Stromal Interaction. Endocrine-Related Cancer, 15, 451-463.
[23] Suzuki, T., Miki, Y., Moriya, T., Akahira, J., Ishida, T., Hirakawa, H., et al. (2007) 5Alpha- Reductase Type 1 and Aromatase in Breast Carcinoma as Regulators of in Situ Androgen Production. International Journal of Cancer, 120, 285-291.
[24] Lukanova, A., Lundin, E., Micheli, A., Arslan, A., Ferrari, P., Rinaldi, S., et al. (2004) Circulating Levels of Sex Steroid Hormones and Risk of Endometrial Cancer in Postmenopausal Women. International Journal of Cancer, 108, 425-432.
[25] Thigpen, J.T., Brady, M.F., Alvarez, R.D., Adelson, M.D., Homesley, H.D., Manetta, A., et al. (1999) Oral Medroxyprogesterone Acetate in the Treatment of Advanced or Recurrent Endometrial Carcinoma: A Dose-Response Study by the Gynecologic Oncology Group. Journal of Clinical Oncology, 17, 1736-1744.
[26] Ushijima, K., Yahata, H., Yoshikawa, H., Konishi, I., Yasugi, T., Saito, T., et al. (2007) Multicenter Phase II Study of Fertility-Sparing Treatment with Medroxyprogesterone Acetate for Endometrial Carcinoma and Atypical Hyperplasia in Young Women. Journal of Clinical Oncology, 25, 2798-2803.
[27] Watanabe, K., Sasano, H., Harada, N., Ozaki, M., Niikura, H., Sato, S., et al. (1995) Aromatase in Human Endometrial Carcinoma and Hyperplasia. Immunohistochemical, in Situ Hybridization, and Biochemical Studies. American Journal of Pathology, 146, 491-500.
[28] Ma, B.B.Y., Oza, A., Eisenhauer, E., Stanimir, G., Carey, M., Chapman, W., et al. (2004) The Activity of Letrozole in Patients with Advanced or Recurrent Endometrial Cancer and Correlation with Biological Markers—A Study of the National Cancer Institute of Canada Clinical Trials Group. International Journal of Gynecological Cancer, 14, 650-658.
[29] Rose, P.G., Brunetto, V.L., VanLe, L., Bell, J., Walker, J.L. and Lee, R.B. (2000) A Phase II Trial of anastrozole in Advanced Recurrent or Persistent Endometrial Carcinoma: A Gynecologic Oncology Group Study. Gynecologic Oncology, 78, 212-216.
[30] Foster, P.A., Chander, S.K., Newman, S.P., Woo, L.W.L., Sutcliffe, O.B., Bubert, C., et al. (2008) A New Therapeutic Strategy against Hormone-Dependent Breast Cancer: The Preclinical Development of a Dual Aromatase and Sulfatase Inhibitor. Clinical Cancer Research, 14, 6469-6477.
[31] Ishida, H., Nakata, T., Suzuki, M., Shiotsu, Y., Tanaka, H., Sato, N., et al. (2007) A Novel Steroidal Selective Steroid Sulfatase Inhibitor KW-2581 Inhibits Sulfated-Estrogen Dependent Growth of Breast Cancer Cells in Vitro and in Animal Models. Breast Cancer Research and Treatment, 106, 215-227.
[32] Ishida, H., Sato, N., Hosogi, J., Tanaka, H. and Kuwabara, T. (2008) Inactivation of Recombinant Human Steroid Sulfatase by KW-2581. Journal of Steroid Biochemistry and Molecular Biology, 108, 17-22.
[33] Purohit, A. and Foster, P. (2012) Steroid Sulfatase Inhibitors for Estrogen- and Androgen-Dependent Cancers. Journal of Endocrinology, 212, 99-110.
[34] Stanway, S.J., Delavault, P., Purohit, A., Woo, L.W.L., Thurieau, C., Potter, B.V.L., et al. (2007) Steroid Sulfatase: A New Target for the Endocrine Therapy of Breast Cancer. Oncologist, 12, 370-374.
[35] Stanway, S.J., Purohit, A., Woo, L.W.L., Sufi, S., Vigushin, D., Ward, R., et al. (2006) Phase I Study of STX 64 (667 Coumate) in Breast Cancer Patients: The First Study of a Steroid Sulfatase Inhibitor. Clinical Cancer Research, 12, 1585-1592.
[36] Audet-Walsh, E., Lépine, J., Grégoire, J., Plante, M., Caron, P., Têtu, B., et al. (2011) Profiling of Endogenous Estrogens, Their Precursors, and Metabolites in Endometrial Cancer Patients: Association with Risk and Relationship to Clinical Characteristics. Journal of Clinical Endocrinology & Metabolism, 96, E330-E339.
[37] Lépine, J., Audet-Walsh, E., Grégoire, J., Têtu, B., Plante, M., Ménard, V., et al. (2010) Circulating Estrogens in Endometrial Cancer Cases and Their Relationship with Tissular Expression of Key Estrogen Biosynthesis and Metabolic Pathways. Journal of Clinical Endocrinology & Metabolism, 95, 2689-2698.
[38] Reed, M.J., Purohit, A., Woo, L.W.L., Newman, S.P. and Potter, B.V.L. (2005) Steroid Sulfatase: Molecular Biology, Regulation, and Inhibition. Endocrine Reviews, 26, 171-202.

comments powered by Disqus

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.