title: Silencing of BRCA2 decreases anoikis and its heterologous expression sensitizes yeast cells to acetic acid-induced programmed cell death.
abstract: Adhesion of normal epithelial cells to the extracellular matrix (ECM) is essential for survival. Cell detachment from ECM induces a specific form of programmed cell death (PCD) termed anoikis. BRCA2, a tumor suppressor gene whose mutations confer predisposition to cancer, has been implicated in the regulation of DNA repair, transcription, cell proliferation, and apoptosis. However, the potential role of BRCA2 in the regulation of anoikis has not been investigated. Here, we found that suppression of BRCA2 expression by short hairpin RNA promoted resistance to anoikis in prostate, breast and thyroid normal epithelial cells, which was accompanied by reduced caspases 3/7 levels and activity. Using yeast as a model, we assessed that expression of human BRCA2 does not induce cell death by itself but it can promote acetic acid-induced PCD (AA-PCD). Induction of BRCA2 expression decreased cell survival and increased the number of cells positive to different apoptotic markers, including DNA fragmentation and phosphatidylserine externalization en route to AA-PCD. A higher increase in ROS levels occurred in the early phase of AA-PCD in BRCA2-expressing yeast cells compared with non-expressing cells. Accordingly, a delay in the initial burst of ROS levels was observed in BRCA2-knockdown anoikis-resistant human cells. Treatment with the antioxidants N-acetylcysteine or ascorbic acid reduced sensitivity to anoikis in human cells and inhibited AA-PCD in yeast cells expressing BRCA2. Taken together, these results show a new function of BRCA2 protein as modulator of anoikis sensitivity through an evolutionarily-conserved molecular mechanism involving regulation of ROS production and/or detoxification by BRCA2 during PCD processes.
authors.affiliation: Institute of Biomembranes and Bioenergetics, National Research Council (CNR), Via Amendola 165/A, 70126, Bari, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Guaragnella
authors.forename: Ersilia
authors.initials: E
authors.lastname: Marra
authors.forename: Alvaro
authors.initials: A
authors.lastname: Galli
authors.forename: Loredana
authors.initials: L
authors.lastname: Moro
authors.forename: Sergio
authors.initials: S
authors.lastname: Giannattasio
mesh_terms: D019342:Acetic Acid / Q000494:pharmacology*; D000111:Acetylcysteine / Q000494:pharmacology; D023102:Anoikis; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D024682:BRCA2 Protein / Q000235:genetics / Q000378:metabolism*; D001940:Breast / Q000166:cytology; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D020868:Gene Silencing; D006801:Humans; D008297:Male; D011467:Prostate / Q000166:cytology; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000378:metabolism; D013961:Thyroid Gland / Q000166:cytology
chemical_list: D000975:Antioxidants; D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D017382:Reactive Oxygen Species; D020169:Caspases; D001205:Ascorbic Acid; D019342:Acetic Acid; D000111:Acetylcysteine