A noncanonical NF-κB pathway through the p50 subunit regulates Bcl-2 overexpression during an oxidative-conditioning hormesis response

doi:10.1016/j.freeradbiomed.2013.04.033

Free Radical Biology and Medicine

Volume 63, October 2013, Pages 41-50

https://doi.org/10.1016/j.freeradbiomed.2013.04.033 Get rights and content

Highlights

•
An oxidative-hormetic model was used to explore Bcl-2 regulation in survival response.
•
Two transductional mechanisms participating in Bcl-2 regulation are proposed.
•
These mechanisms converge in activating the nuclear transcription factor NF-κB.
•
The noncanonical p50 subunit participates during the oxidative-hormetic response.

Abstract

Cells can respond to damage and stress by activating various repair and survival pathways. One of these responses can be induced by preconditioning the cells with sublethal stress to provoke a prosurvival response that will prevent damage and death, and which is known as hormesis. Bcl-2, an antiapoptotic protein recognized by its antioxidant and prosurvival functions, has been documented to play an important role during oxidative-conditioning hormesis. Using an oxidative-hormetic model, which was previously established in the L929 cell line by subjecting the cells to a mild oxidative stress of 50 μM H₂O₂ for 9 h, we identified two different transductional mechanisms that participate in the regulation of Bcl-2 expression during the hormetic response. These mechanisms converge in activating the nuclear transcription factor NF-κB. Interestingly, the noncanonical p50 subunit of the NF-κB family is apparently the subunit that participates during the oxidative-hormetic response.

Graphical abstract

Section snippets

Chemicals

All chemicals and reagents were of the highest analytical grade and most of them were purchased from Sigma (St. Louis, MO, USA). Reagents obtained from other suppliers are detailed in the text.

Cell culture

Mouse L929 lung fibroblasts were cultured at 37 °C in an atmosphere of 95% air and 5% CO₂ as described elsewhere [37].

Cellular viability and oxidative-conditioning hormesis

L929 cells were seeded at 1×10⁵ cells/well into 24-well plates (Corning, Acton, MA, USA) and were treated with 50 μM H₂O₂ for 9 h, to induce the OCH response, and it was compared in some

PI3K, Akt, and PKC regulate Bcl-2 expression during the hormetic response

Previously we reported the role played by Bcl-2 during the hormetic response induced by mild oxidative stress [36]. To determine the signaling mechanism that regulates Bcl-2 expression, here we evaluated the participation of the main kinases known for their contribution to cell survival against oxidative stress.

PI3K, Akt, and PKC-α phosphorylation was monitored at short time points after OCH treatment (50 µM H₂O₂) (Fig. 1). When PI3K and p-PI3K were evaluated, a fast activation was observed by

Discussion

In this work we have established two different signaling mechanisms that participate in the regulation of Bcl-2 expression during the hormetic response. The hormetic model or OCH was previously established in the L929 cell line [36], subjecting the cells to a mild oxidative stress (50 μM H₂O₂) for 9 h.

The first mechanism involves PI3K, Akt, NF-κB, and Bcl-2 and has been described before as part of the survival and antioxidant response against several toxic stimuli in various cell types [27], [39]

Acknowledgments

The authors thank Dr. A. Hernández from CINVESTAV for generously donating to us the actin antibody and M.Sc. R. Lazzarini and the CBS–UAMI Confocal Core for confocal images acquisition and analysis. This work was supported by CONACyT Grants CB-2006-1-59659 and CB-2012-1-178349 as well as the “Red Temática de Investigación en Salud y Desarrollo Social” from CONACyT and INGER: DI-PI004/2012.

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Original ContributionA noncanonical NF-κB pathway through the p50 subunit regulates Bcl-2 overexpression during an oxidative-conditioning hormesis response

Highlights

Abstract

Graphical abstract

Section snippets

Chemicals

Cell culture

Cellular viability and oxidative-conditioning hormesis

PI3K, Akt, and PKC regulate Bcl-2 expression during the hormetic response

Discussion

Acknowledgments

Toxicol. Appl. Pharmacol.

Ageing Res. Rev.

Brain Res.

Brain Res. Bull.

Food Chem. Toxicol.

Eur. J. Pharmacol

Semin. Cancer Biol.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

FEBS Lett

Toxicol. Appl. Pharmacol

Cell. Signalling

Mol. Cell

Free Radic. Biol. Med.

FEBS Lett

J. Biol. Chem.

Anal. Biochem.

Trends Pharmacol. Sci.

Free Radic. Biol. Med.

Cancer Lett.

Free Radic. Biol. Med.

Brain Res.

FEBS Lett.

Am. J. Pathol.

Exp. Eye Res.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

A perspective on the scientific, philosophical, and policy dimensions of hormesis

Dose–Response

p27(Kip1) inhibits systemic autoimmunity through the control of Treg cell activity and differentiation

Arthritis Rheum.

Simultaneously targeting Bcl-2 and Akt pathways sensitizes nasopharyngeal carcinoma to tumor necrosis factor-related apoptosis-inducing ligand

Cancer Biother. Radiopharm.

Borneol alleviates oxidative stress via upregulation of Nrf2 and Bcl-2 in SH-SY5Y cells

Pharm. Biol.

α-Mangostin induces apoptosis in human chondrosarcoma cells through downregulation of ERK/JNK and Akt signaling pathway

Agric. Food Chem

Rapid reactive oxygen species (ROS) generation induced by curcumin leads to caspase-dependent and -independent apoptosis in L929 cells

Free Radic. Biol. Med.

Tea polyphenols induce apoptosis through mitochondrial pathway and by inhibiting nuclear factor-kappaB and Akt activation in human cervical cancer cells

Oncol. Res.

Original Contribution
A noncanonical NF-κB pathway through the p50 subunit regulates Bcl-2 overexpression during an oxidative-conditioning hormesis response