Research Paper Volume 11, Issue 22 pp 10532—10556

Distinct effects of epirubicin, cisplatin and cyclophosphamide on ovarian somatic cells of prepuberal ovaries

Serena Marcozzi 1, , Valerio Rossi 1, , Giulia Salvatore 1, , Francesca Di Rella 2, , Massimo De Felici 1, , Francesca Gioia Klinger 1, ,

  • 1 Department of Biomedicine and Prevention, Section of Histology and Embryology, Faculty of Medicine and Surgery, University of Rome Tor Vergata, Rome, Italy
  • 2 Medical Oncology, Department of Senology, National Cancer Institute, IRCCS Foundation G. Pascale, Naples, Italy

received: August 6, 2019 ; accepted: November 8, 2019 ; published: November 11, 2019 ;

https://doi.org/10.18632/aging.102476
How to Cite

Copyright © 2019 Marcozzi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

In vitro culture models were used to characterize the effects of chemotherapeutic drugs and of LH on somatic cells from prepuberal mouse ovaries. All cell types (pre- and granulosa cells, pre-thecal and OSE cells) underwent apoptosis following Epirubicin (0.5μM) exposure for 24hrs (about 60%) and 48hrs (>80%). Cisplatin (10μM) and the Cyclophosphamide active metabolite, Phosphoramide Mustard (10μM), didn’t cause apoptosis in 90% of pre-thecal and pre-granulosa cells up to 72hrs of exposure, although they suffered extensive DNA damage and cell cycle arrest, and acquired stress induced premature senescence (SIPS) features. Cultured granulosa cells didn’t show evident DNA damage and remained viable without acquiring SIPS features; OSE cells were resistant to apoptosis and SIPS but not to DNA damage. These latter, like pre-thecal and pre-granulosa cells, were able of efficient DNA repair involving MLH1-dependent MMR pathways. SIPS features were also observed in ovary after in vivo treatment with Cisplatin. LH (200mIU/mL) didn’t significantly influence apoptosis, SIPS and DNA damage but favoured DNA repair. These results show that somatic cells of prepuberal ovary response to drugs in different ways, either undergoing apoptosis or SIPS, either showing resistance to Cisplatin and Phosphoramide Mustard. Moreover, a new role of LH in promoting DNA repair was shown.

Abbreviations

AMH: Anti-müllerian hormone; CPM: Cyclophosphamide; CS: Cisplatin; CYP17A1: Cytochrome P450 17A1; DDR: DNA damage response; dpp: Days postpartum; EOC: Epithelial ovarian cancer; EPI: Epirubicin; ERCC1: Excision Repair 1; FOXL2: Forkhead box L2; GADPH: Glyceraldehyde 3-phosphate dehydrogenase; GCs: Granulosa cells; Gli1: Glioma-Associated Oncogene Homolog 1; HR: Homologous Recombination; LH: Luteinizing hormone; LHCGR: Luteinizing hormone/Choriogonadotropin receptor; MLH1: MutL homolog 1; MMR: DNA Mismatch repair; NER: Nucleotide excision repair; OSE: Ovarian surface epithelium; pan-CKs: pan cytokeratin; pGCs: Precursor granulosa cells; PI: Propidium Iodide; PM: Phosphoramide mustard; PMF: primordial follicle; POI: Premature ovarian insufficiency; POs: Primordial follicle-enclosed oocytes (POs); RAD51: RAD51 Recombinase; SASP: senescence-associated secretory phenotype; SIPS: stress induced premature senescence; TCs: Follicular theca cells; αSMA: Alpha-smooth muscle actin; γH2AX: phosphorylated form of H2A histone family member X.