In vitro antitumor activity of progesterone in human adrenocortical carcinoma
The management of patients with adrenocortical carcinoma (ACC) is challenging. As mitotane and chemotherapy show limited efficacy, there is an urgent need to develop therapeutic approaches. The aim of this study was to investigate the antitumor activity of progesterone and explore the molecular mechanisms underlying its cytotoxic effects in the NCI-H295R cell line and primary cell cultures derived from ACC patients.
Cell viability, cell cycle, and apoptosis were analyzed in untreated and progesterone-treated ACC cells. The ability of progesterone to affect the Wnt/β-catenin pathway in NCI-H295R cells was investigated by immunofluorescence. Progesterone and mitotane combination experiments were also performed to evaluate their interaction on NCI-H295R cell viability.
We demonstrated that progesterone exerted a concentration-dependent inhibition of ACC cell viability. Apoptosis was the main mechanism, as demonstrated by a significant increase of apoptosis and cleaved-Caspase-3 levels. Reduction of β-catenin nuclear translocation may contribute to the progesterone cytotoxic effect. The progesterone antineoplastic activity was synergically increased when mitotane was added to the cell culture medium.
Our results show that progesterone has antineoplastic activity in ACC cells. The synergistic cytotoxic activity of progesterone with mitotane provides the rationale for testing this combination in a clinical study.
KeywordsAdrenocortical carcinoma Progesterone Progesterone receptor Cell viability
Interval of Confidence
progesterone membrane receptor
progesterone receptor membrane component 1
This work was supported by: AIRC project IG17678 (PI: M.T.); AIRC project IG14411 (PI: A.B.); Fondazione Camillo Golgi, Brescia; University of Brescia local grants; private donation of “gli Amici di Andrea” in memory of Andrea Gadeschi; private grant from the amateur dramatics group “Attori non per caso”, Parish church of Collio Valtrompia (Brescia). M.F. was supported by a grant from the Italian Society of Pharmacology.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 5.A. Berruti, M. Terzolo, P. Sperone, A. Pia, S. Della Casa, D.J. Gross, C. Carnaghi, P. Casali, F. Porpiglia, F. Mantero, G. Reimondo, A. Angeli, L. Dogliotti, Etoposide, doxorubicin and cisplatin plus mitotane in the treatment of advanced adrenocortical carcinoma: a large prospective phase II trial. Endocr. Relat. Cancer 12, 657–666 (2005)CrossRefGoogle Scholar
- 6.A. Berruti, M. Fassnacht, H. Haak, T. Else, E. Baudin, P. Sperone, M. Kroiss, T. Kerkhofs, A.R. Williams, A. Ardito, S. Leboulleux, M. Volante, T. Deutschbein, R. Feelders, C. Ronchi, S. Grisanti, H. Gelderblom, F. Porpiglia, M. Papotti, G.D. Hammer, B. Allolio, M. Terzolo, Prognostic role of overt hypercortisolism in completely operated patients with adrenocortical cancer. Eur. Urol. 65, 832–838 (2014)CrossRefGoogle Scholar
- 7.R. Libé, I. Borget, C.L. Ronchi, B. Zaggia, M. Kroiss, T. Kerkhofs, J. Bertherat, M. Volante, M. Quinkler, O. Chabre, M. Tabarin, F. Beuschlein et al.. Prognostic factors in stage III-IV adrenocortical carcinomas (ACC): an European Network for the Study of Adrenal Tumor (ENSAT) study. Ann. Oncol. 26, 2119–2125 (2015)CrossRefGoogle Scholar
- 9.S. Puglisi, P. Perotti, D. Cosentini, E. Roca, V. Basile, A. Berruti, M. Terzolo, Decision-making for adrenocortical carcinoma: surgical, systemic, and endocrine management options. Expert. Rev. AntiCancer Ther. 8(11), 1–9 (2018)Google Scholar
- 10.C. Fiorentini, M. Fragni, P. Perego, S. Vezzoli, S.A. Bonini, M. Tortoreto, D. Galli, M. Claps, G.A. Tiberio, M. Terzolo, C. Missale, M. Memo, G. Procopio, N. Zaffaroni, A. Berruti, S. Sigala, Antisecretive and antitumor activity of abiraterone acetate in human adrenocortical cancer: a preclinical study. J. Clin. Endocrinol. Metab. 101, 4594–4602 (2016)CrossRefGoogle Scholar
- 11.G. Attard, A.H. Reid, R.J. Auchus, B.A. Hughes, A.M. Cassidy, E. Thompson, N.B. Oommen, E. Folkerd, M. Dowsett, W. Arlt, J.S. de Bono, Clinical and biochemical consequences of CYP17A1 inhibition with abiraterone given with and without exogenous glucocorticoids in castrate men with advanced prostate cancer. J. Clin. Endocrinol. Metab. 97, 507–516 (2012)CrossRefGoogle Scholar
- 24.C. Fiorentini, S. Bodei, F. Bedussi, M. Fragni, S.A. Bonini, C. Simeone, D. Zani, A. Berruti, C. Missale, M. Memo, P.F. Spano, S. Sigala, GPNMB/OA protein increases the invasiveness of human metastatic prostate cancer cell lines DU145 and PC3 through MMP-2 and MMP-9 activity. Exp. Cell Res. 323, 100–111 (2014)CrossRefGoogle Scholar
- 25.V. Porrini, I. Sarnico, M. Benarese, C. Branca, M. Mota, A. Lanzillotta, A. Bellucci, E. Parrella, L. Faggi, P.F. Spano, B.P. Imbimbo, M. Pizzi, Neuroprotective and anti-apoptotic efects of CSP-1103 in primary cortical neurons exposed to oxygen and glucose deprivation. Int. J. Mol. Sci. 18, E184 (2017)CrossRefGoogle Scholar
- 29.A. Chimento, R. Sirianni, I. Casaburi, F. Zolea, P. Rizza, P. Avena, R. Malivindi, A. De Luca, C. Campana, E. Martire, F. Domanico, F. Fallo, G. Carpinelli, L. Cerquetti, D. Amendola, A. Stigliano, V. Pezzi, GPER agonist G-1 decreases adrenocortical carcinoma (ACC) cell growth in vitro and in vivo. Oncotarget 6, 19190–19203 (2015)CrossRefGoogle Scholar
- 30.R.S.Y. Wong, Apoptosis in cancer: from pathogenesis to treatment. J. Exp. Clin. Cancer Res. 30, 87 (2011)Google Scholar
- 35.A. Leibovitz, W.M. McCombs 3rd, D. Johnston, C.E. McCoy, J.C. Stinson, New human cancer cell culture lines. I. SW-13, small-cell carcinoma of the adrenal cortex. J. Natl. Cancer Inst. 51, 691–697 (1973)Google Scholar
- 40.K. Horita, N. Inase, S. Miyake, B. Formby, H. Toyoda, Y. Yoshizawa, Progesterone induces apoptosis in malignant mesothelioma cells. Anticancer Res. 21, 3871–3874 (2001)Google Scholar
- 42.Y. Wang, P. Hanifi-Moghaddam, E.E. Hanekamp, H.J. Kloosterboer, P. Franken, J. Veldscholte, H.C. van Doorn, P.C. Ewing, J.J. Kim, J.A. Grootegoed, C.W. Burger, R. Fodde, L.J. Blok, Progesterone inhibition of Wnt/beta-catenin signaling in normal endometrium and endometrial cancer. Clin. Cancer Res. 15, 5784–5793 (2009)CrossRefGoogle Scholar