DNA repair protein DNA-PK protects PC12 cells from oxidative stress-induced apoptosis involving AKT phosphorylation.

Autor: Cardinale A; Molecular and Cellular Neurobiology, IRCCS San Raffaele Roma, Via di Val Cannuta 247, 00166, Rome, Italy., Saladini S; Molecular and Cellular Neurobiology, IRCCS San Raffaele Roma, Via di Val Cannuta 247, 00166, Rome, Italy., Lupacchini L; Molecular and Cellular Neurobiology, IRCCS San Raffaele Roma, Via di Val Cannuta 247, 00166, Rome, Italy., Ruspantini I; FAST. Istituto Superiore di Sanita', Viale Regina Elena 299, 00161, Rome, Italy., De Dominicis C; Molecular and Cellular Neurobiology, IRCCS San Raffaele Roma, Via di Val Cannuta 247, 00166, Rome, Italy.; Department of Neuroscience, Istituto Superiore di Sanita', Viale Regina Elena 299, 00161, Rome, Italy., Papale M; Molecular and Cellular Neurobiology, IRCCS San Raffaele Roma, Via di Val Cannuta 247, 00166, Rome, Italy., Silvagno F; Department of Oncology, University Torino, via Santena 5 bis, 10126, Torino, Italy., Garaci E; University San Raffaele, Via di Val Cannuta 247, 00166, Rome, Italy., Mollinari C; Department of Neuroscience, Istituto Superiore di Sanita', Viale Regina Elena 299, 00161, Rome, Italy.; Institute of Translational Pharmacology, National Research Council, Via Fosso del Cavaliere 100, 00133, Rome, Italy., Merlo D; Department of Neuroscience, Istituto Superiore di Sanita', Viale Regina Elena 299, 00161, Rome, Italy. daniela.merlo@iss.it.
Jazyk: angličtina
Zdroj: Molecular biology reports [Mol Biol Rep] 2022 Feb; Vol. 49 (2), pp. 1089-1101. Date of Electronic Publication: 2021 Nov 19.
DOI: 10.1007/s11033-021-06934-5
Abstrakt: Background: Emerging evidence suggest that DNA-PK complex plays a role in the cellular response to oxidative stress, in addition to its function of double strand break (DSB) repair. In this study we evaluated whether DNA-PK participates in oxidative stress response and whether this role is independent of its function in DNA repair.
Methods and Results: We used a model of H 2 O 2 -induced DNA damage in PC12 cells (rat pheochromocytoma), a well-known neuronal tumor cell line. We found that H 2 O 2 treatment of PC12 cells induces an increase in DNA-PK protein complex levels, along with an elevation of DNA damage, measured both by the formation of γΗ2ΑX foci, detected by immunofluorescence, and γH2AX levels detected by western blot analysis. After 24 h of cell recovery, γΗ2ΑX foci are repaired both in the absence and presence of DNA-PK kinase inhibitor NU7026, while an increase of apoptotic cells is observed when DNA-PK activity is inhibited, as revealed by counting pycnotic nuclei and confirmed by FACS analysis. Our results suggest a role of DNA-PK as an anti-apoptotic factor in proliferating PC12 cells under oxidative stress conditions. The anti-apoptotic role of DNA-PK is associated with AKT phosphorylation in Ser473. On the contrary, in differentiated PC12 cells, were the main pathway to repair DSBs is DNA-PK-mediated, the inhibition of DNA-PK activity causes an accumulation of DNA damage.
Conclusions: Taken together, our results show that DNA-PK can protect cells from oxidative stress induced-apoptosis independently from its function of DSB repair enzyme.
(© 2021. The Author(s).)
Databáze: MEDLINE
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