Preclinical Evidence of Progesterone as a New Pharmacological Strategy in Human Adrenocortical Carcinoma Cell Lines

Abstract

Background: Adrenocortical cancer (ACC) is a rare malignancy with a dismal prognosis. The treatment includes mitotane and EDP chemotherapy (etoposide, doxorubicin, and cisplatin). However, new therapeutic approaches for advanced ACC are needed, particularly targeting the metastatic process. Here, we deepen the role of progesterone as a new potential drug for ACC, in line with its antitumoral effect in other cancers. Methods: NCI-H295R, MUC-1, and TVBF-7 cell lines were used and xenografted in zebrafish embryos. Migration and invasion were studied using transwell assays, and MMP2 activity was studied using zymography. Apoptosis and cell cycle were analyzed by flow cytometry. Results: Progesterone significantly reduced xenograft tumor area and metastasis formation in embryos injected with metastatic lines, MUC-1 and TVBF-7. These results were confirmed in vitro, where the reduction of invasion was mediated, at least in part, by the decrease in MMP2 levels. Progesterone exerted a long-lasting effect in metastaticcells. Progesterone caused apoptosis in NCI-H295R and MUC-1, inducing changes in the cell-cycle distribution, while autophagy was predominantly activated in TVBF-7 cells. Conclusion: Our results give support to the role of progesterone in ACC. The involvement of its analog (megestrol acetate) in reducing ACC progression in ACC patients undergoing EDP-M therapy is now under investigation in the PESETA phase II clinical study.

Feasibility and Activity of Megestrol Acetate in Addition to Etoposide, Doxorubicin, Cisplatin, and Mitotane as First-Line Therapy in Patients with Metastatic/Unresectable Adrenocortical Carcinoma with Low Performance Status

“…More recently, the antineoplastic effect of progesterone on the above-cited cell lines was confirmed in an in vivo zebrafish embryo xenograft model. In this model, progesterone was also found to inhibit metastasis formation in tumor xenografts derived from MUC-1 and TVBF-7 metastatic cells, confirming the in vitro results where migration and invasion abilities were significantly suppressed by this hormone [ 24 ].…”

“…Since progestins are efficacious in counteracting cancer-associated anorexia and cachexia, we administered megestrol acetate in association with EDP-M in ACC patients with compromised PS with the aim to improve the patient’s general conditions and mitigate some side effects of EDP-M. In addition, based on preclinical data from our group showing a specific antineoplastic activity of progesterone in in vitro and in vivo ACC experimental models [ 19 , 20 , 23 , 24 ] and a synergistic effect when combined with other cytotoxic therapies, we also explored the impact of the addition of megestrol acetate on the safety of the EDP-M regimen. It should be underlined, however, that in in vitro experimental models, synthetic progestin megestrol acetate is able to bind and activate other steroid hormone receptors, including glucocorticoid receptors [ 31 , 32 , 33 ].…”

“…Metastasis-derived cell models, namely MUC-1 and TVBF-7 cell lines, were found to be less sensitive to the Pg effect due to weaker PgR expression compared to NCI-H295R cells [ 23 ]. Interestingly, the cytotoxic effect of progesterone, when present, is maintained after drug withdrawal [ 24 ].…”

The CDK Inhibitor Dinaciclib Improves Cisplatin Response in Nonseminomatous Testicular Cancer: A Preclinical Study

“…After 3 days (T3), the drug effects were observed. To evaluate the effect of dinaciclib on tumor growth, Tg (kdrl:EGFP) zebrafish embryos at 48 hpf were dechorionated, anesthetized with 0.042 mg/mL tricaine and microinjected with labeled CP-sensitive/resistant NT2/D1 and CP-sensitive/resistant NCCIT cells into the subperidermal space of the yolk sac as described [ 29 , 30 ]. Approximately 250 cells/4 nL were injected into each embryo (about 50 embryos/group), and embryos were maintained in PTU/fish water in a 32 °C incubator to allow tumor cell growth.…”