title: BNC2 is a putative tumor suppressor gene in high-grade serous ovarian carcinoma and impacts cell survival after oxidative stress.
abstract: Rs3814113 is the single-nucleotide polymorphism (SNP) showing the strongest association with high-grade serous ovarian carcinoma (HGSOC) incidence and is located in an intergenic region about 44 kb downstream of basonuclin 2 (BNC2) gene. Lifetime number of ovulations is associated with increased risk to develop HGSOC, probably because of cell damage of extrauterine Müllerian epithelium by ovulation-induced oxidative stress. However, the impact of low-penetrance HGSOC risk alleles (e.g. rs3814113) on the damage induced by oxidative stress remains unclear. Therefore, the purpose of this study was to investigate whether rs3814113 genetic interval regulates BNC2 expression and whether BNC2 expression levels impact on cell survival after oxidative stress. To do this, we analyzed gene expression levels of BNC2 first in HGSOC data sets and then in an isogenic cell line that we engineered to carry a 5 kb deletion around rs3814113. Finally, we silenced BNC2 and measured surviving cells after hydrogen peroxide (H2O2) treatment to simulate oxidative stress after ovulation. In this paper, we describe that BNC2 expression levels are reduced in HGSOC samples compared with control samples, and that BNC2 expression levels decrease following oxidative stress and ovulation in vitro and in vivo, respectively. Moreover, deletion of 5 kb surrounding rs3814113 decreases BNC2 expression levels in an isogenic cell line, and silencing of BNC2 expression levels increases cell survival after H2O2treatment. Altogether, our findings suggest that the intergenic region located around rs3814113 regulates BNC2 expression, which in turn affects cell survival after oxidative stress response. Indeed, HGSOC samples present lower BNC2 expression levels that probably, in the initial phases of oncogenic transformation, conferred resistance to oxidative stress and ultimately reduced the clearance of cells with oxidative-induced damages.
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Cesaratto
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Grisard
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.|Department of Life and Reproduction Sciences, University of Verona, Verona, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Coan
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Zandonà
authors.affiliation: Division of Experimental and Clinical Pharmacology, Department of Translational Research Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: De Mattia
authors.affiliation: Department of Oncology, University Hospital of Udine, Udine, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Poletto
authors.affiliation: Division of Experimental and Clinical Pharmacology, Department of Translational Research Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Erika
authors.initials: E
authors.lastname: Cecchin
authors.affiliation: Department of Oncology, University Hospital of Udine, Udine, Italy.|Department of Medical and Biological Sciences, University of Udine, Udine, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Puglisi
authors.affiliation: Division of Pathology, Department of Translational Research, CRO Aviano National Cancer Institute, Aviano, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Canzonieri
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Mucignat
authors.affiliation: Clinical and Experimental Onco-Hematology Unit, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Zucchetto
authors.affiliation: Department of Life Sciences, University of Trieste, Trieste, Italy.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Stocco
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Colombatti
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Milena S
authors.initials: MS
authors.lastname: Nicoloso
authors.affiliation: Division of Experimental Oncology2, Department of Translational Research, Centro di Riferimento Oncologico (CRO Aviano), National Cancer Institute, Aviano, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Spizzo
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D018284:Cystadenocarcinoma, Serous / Q000235:genetics* / Q000473:pathology*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D016147:Genes, Tumor Suppressor; D008040:Genetic Linkage; D015894:Genome, Human; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D051379:Mice; D060787:Neoplasm Grading; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology*; D018384:Oxidative Stress / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics
chemical_list: C484527:BNC2 protein, human; D004268:DNA-Binding Proteins; D006861:Hydrogen Peroxide