Acute Valproate Exposure Induces Mitochondrial Biogenesis and Autophagy with FOXO3a Modulation in SH-SY5Y Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Cytotoxicity Test
2.3. LDH Assay
2.4. ATP Lite Assay
2.5. Mitochondrial Membrane Potential Assay
2.6. Oxidative Stress Analysis
2.7. Mitochondrial (mt) DNA Copy Number Assay
2.8. Western Blotting
2.9. Immunocytochemistry
2.10. Immunoprecipitation
2.11. Autophagy Detection
2.12. Statistical Analysis
3. Results
3.1. VPA Induced Cytotoxicity with Changes to the Mitochondria Membrane Potential and ATP Production in the SH-SY5Y Cell Line
3.2. Change of mtDNA Copy Numbers and Protein Levels for Mitochondria Biogenesis-Related Genes with VPA Acute Exposure in the SH-SY5Y Cell Line
3.3. Acute VPA Treatment Induced FOXO3a Expression in SY-SY5Y Cells
3.4. VPA Modulates the FOXO3a Acetylation with Changes to SIRT1 and SIRT3 Protein Levels in SH-SY5Y Cells
3.5. VPA Treatment Induced Autophagy in SH-SY5Y Cells
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
- Khurana, D.S.; Valencia, I.; Goldenthal, M.J.; Legido, A. Mitochondrial dysfunction in epilepsy. Semin. Pediatr. Neurol. 2013, 20, 176–187. [Google Scholar] [CrossRef] [PubMed]
- Komulainen, T.; Lodge, T.; Hinttala, R.; Bolszak, M.; Pietilä, M.; Koivunen, P.; Hakkola, J.; Poulton, J.; Morten, K.J.; Uusimaa, J. Sodium valproate induces mitochondrial respiration dysfunction in HepG2 in vitro cell model. Toxicology 2015, 331, 47–56. [Google Scholar] [CrossRef] [PubMed]
- Terranova-Barberio, M.; Roca, M.S.; Zotti, A.I.; Leone, A.; Bruzzese, F.; Vitagliano, C.; Scogliamiglio, G.; Russo, D.; D’Angelo, G.; Franco, R.; et al. Valproic acid potentiates the anticancer activity of capecitabine in vitro and in vivo in breast cancer models via induction of thymidine phosphorylase expression. Oncotarget 2016, 7, 7715–7731. [Google Scholar] [CrossRef] [PubMed]
- Duenas-Gonzalez, A.; Candelaria, M.; Perez-Plascencia, C.; Perez-Cardenas, E.; de la Cruz-Hernandez, E.; Herrera, L.A. Valproic acid as epigenetic cancer drug: Preclinical, clinical and transcriptional effects on solid tumors. Cancer Treat. Rev. 2008, 34, 206–222. [Google Scholar] [CrossRef]
- Xia, Q.; Zheng, Y.; Jiang, W.; Huang, Z.; Wang, M.; Rodriguez, R.; Jin, X. Valproic acid induces autophagy by suppressing the Akt/mTOR pathway in human prostate cancer cells. Oncol. Lett. 2016, 12, 1826–1832. [Google Scholar] [CrossRef] [Green Version]
- Wang, Y.; Hao, C.L.; Zhang, Z.H.; Wang, L.H.; Yan, L.N.; Zhang, R.J.; Lin, L.; Yang, Y. Valproic Acid Increased Autophagic Flux in human Multiple Myeloma Cells in Vitro. Biomed. Pharmacother. 2020, 127, 110167. [Google Scholar] [CrossRef]
- Cheng, Z. The FoxO-Autophagy Axis in Health and Disease. Trends Endocrin. Met. 2019, 30, 658–671. [Google Scholar] [CrossRef] [PubMed]
- Brown, A.K.; Webb, A.E. Regulation of FOXO Factors in Mammalian Cells. Curr. Top. Dev. Biol. 2018, 127, 165–192. [Google Scholar] [CrossRef] [PubMed]
- Willcox, B.J.; Donlon, T.A.; He, Q.; Chen, R.; Grove, J.S.; Yano, K.; Masaki, K.H.; Willcox, D.C.; Rodriguez, B.; Curb, J.D. FOXO3A genotype is strongly associated with human longevity. Proc. Natl. Acad. Sci. USA 2008, 105, 13987–13992. [Google Scholar] [CrossRef] [Green Version]
- Santo, E.E.; Stroeken, P.; Sluis, P.V.; Koster, J.; Versteeg, R.; Westerhout, E.M. FOXO3a is a major target of inactivation by PI3K/AKT signaling in aggressive neuroblastoma. Cancer Res. 2013, 73, 2189–2198. [Google Scholar] [CrossRef] [Green Version]
- Qian, Z.; Ren, L.; Wu, D.; Yang, X.; Zhou, Z.; Nie, Q.; Jiang, G.; Xue, S.; Weng, W.; Qiu, Y.; et al. Overexpression of FoxO3a is associated with glioblastoma progression and predicts poor patient prognosis. Int. J. Cancer 2017, 140, 2792–2804. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Audesse, A.J.; Dhakal, S.; Hassell, L.A.; Gardell, Z.; Nemtsova, Y.; Webb, A.E. FOXO3 directly regulates an autophagy network to functionally regulate proteostasis in adult neural stem cells. PLoS Genet. 2019, 15, e1008097. [Google Scholar] [CrossRef]
- Lim, S.W.; Jin, L.; Luo, K.; Jin, J.; Shin, Y.J.; Hong, S.Y.; Yang, C.W. Klotho enhances FoxO3-mediated manganese superoxide dismutase expression by negatively regulating PI3K/AKT pathway during tacrolimus-induced oxidative stress. Cell Death Dis. 2017, 8, e2972. [Google Scholar] [CrossRef] [PubMed]
- Grossi, V.; Fasano, C.; Celestini, V.; Lepore Signorile, M.; Sanese, P.; Simone, C. Chasing the FOXO3: Insights into Its New Mitochondrial Lair in Colorectal Cancer Landscape. Cancers 2019, 11, 414. [Google Scholar] [CrossRef] [Green Version]
- Khalil, M.A.; Hraběta, J.; Groh, T.; Procházka, P.; Doktorová, H.; Eckschlager, T. Valproic Acid Increases CD133 Positive Cells that Show Low Sensitivity to Cytostatics in Neuroblastoma. PLoS ONE 2016, 11, e0162916. [Google Scholar] [CrossRef]
- Gu, S.; Tian, Y.; Chlenski, A.; Salwen, H.R.; Lu, Z.; Raj, J.U.; Yang, Q. Valproic acid shows a potent antitumor effect with alteration of DNA methylation in neuroblastoma. Anti-Cancer Drug 2012, 23, 1054–1066. [Google Scholar] [CrossRef] [Green Version]
- Blaheta, R.A.; Michaelis, M.; Natsheh, I.; Hasenberg, C.; Weich, E.; Relja, B.; Jonas, D.; Doerr, H.W.; Cinatl, J., Jr. Valproic acid inhibits adhesion of vincristine- and cisplatin-resistant neuroblastoma tumour cells to endothelium. Br. J. Cancer 2007, 96, 1699–1706. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001, 25, 402–408. [Google Scholar] [CrossRef]
- Liu, Y.; Ao, X.; Ding, W.; Ponnusamy, M.; Wu, W.; Hao, X.; Yu, W.; Wang, Y.; Li, P.; Wang, J. Critical role of FOXO3a in carcinogenesis. Mol. Cancer 2018, 17, 104. [Google Scholar] [CrossRef] [Green Version]
- Shukla, S.; Bhaskaran, N.; Maclennan, G.T.; Gupta, S. Deregulation of FoxO3a accelerates prostate cancer progression in TRAMP mice. Prostate 2013, 73, 1507–1517. [Google Scholar] [CrossRef] [Green Version]
- Shoeb, M.; Ramana, K.V.; Srivastava, S.K. Aldose reductase inhibition enhances TRAIL-induced human colon cancer cell apoptosis through AKT/FOXO3a-dependent upregulation of death receptors. Free Radic. Biol. Med. 2013, 63, 280–290. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lützner, N.; Kalbacher, H.; Krones-Herzig, A.; Rösl, F. FOXO3 is a glucocorticoid receptor target and regulates LKB1 and its own expression based on cellular AMP levels via a positive autoregulatory loop. PLoS ONE 2012, 7, e42166. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ryu, H.; Lee, J.; Olofsson, B.A.; Mwidau, A.; Dedeoglu, A.; Escudero, M.; Flemington, E.; Azizkhan-Clifford, J.; Ferrante, R.J.; Ratan, R.R. Histone deacetylase inhibitors prevent oxidative neuronal death independent of expanded polyglutamine repeats via an Sp1-dependent pathway. Proc. Natl. Acad. Sci. USA 2003, 100, 4281–4286. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yu, Y.; Peng, K.; Li, H.; Zhuang, R.; Wang, Y.; Li, W.; Yu, S.; Liang, L.; Xu, X.; Liu, T. SP1 upregulated FoxO3a promotes tumor progression in colorectal cancer. Oncol. Rep. 2018, 39, 2235–2242. [Google Scholar] [CrossRef] [PubMed]
- Kannike, K.; Sepp, M.; Zuccato, C.; Cattaneo, E.; Timmusk, T. Forkhead transcription factor FOXO3a levels are increased in Huntington disease because of overactivated positive autofeedback loop. J. Biol. Chem. 2014, 289, 32845–32857. [Google Scholar] [CrossRef] [Green Version]
- Kops, G.J.; Dansen, T.B.; Polderman, P.E.; Saarloos, I.; Wirtz, K.W.; Coffer, P.J.; Huang, T.T.; Bos, J.L.; Medema, R.H.; Burgering, B.M. Forkhead transcription factor FOXO3a protects quiescent cells from oxidative stress. Nature 2002, 419, 316–321. [Google Scholar] [CrossRef]
- Tseng, A.H.; Shieh, S.S.; Wang, D.L. SIRT3 deacetylates FOXO3 to protect mitochondria against oxidative damage. Free Radic. Biol. Med. 2013, 63, 222–234. [Google Scholar] [CrossRef]
- Vernier, M.; Dufour, C.R.; McGuirk, S.; Scholtes, C.; Li, X.; Bourmeau, G.; Kuasne, H.; Park, M.; St-Pierre, J.; Audet-Walsh, E.; et al. Estrogen-related receptors are targetable ROS sensors. Gene Dev. 2020, 34, 544–559. [Google Scholar] [CrossRef]
- Salminen, A.; Kaarniranta, K.; Kauppinen, A. Crosstalk between Oxidative Stress and SIRT1: Impact on the Aging Process. Int. J. Mol. Sci. 2013, 14, 3834–3859. [Google Scholar] [CrossRef] [Green Version]
- Chen, Z.; Shentu, T.P.; Wen, L.; Johnson, D.A.; Shyy, J.Y. Regulation of SIRT1 by oxidative stress-responsive miRNAs and a systematic approach to identify its role in the endothelium. Antioxid. Redox Sign. 2013, 19, 1522–1538. [Google Scholar] [CrossRef] [Green Version]
- Caito, S.; Rajendrasozhan, S.; Cook, S.; Chung, S.; Yao, H.; Friedman, A.E.; Brookes, P.S.; Rahman, I. SIRT1 is a redox-sensitive deacetylase that is post-translationally modified by oxidants and carbonyl stress. FASEB J. 2010, 24, 3145–3159. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Cai, W.; Ramdas, M.; Zhu, L.; Chen, X.; Striker, G.E.; Vlassara, H. Oral advanced glycation endproducts (AGEs) promote insulin resistance and diabetes by depleting the antioxidant defenses AGE receptor-1 and sirtuin 1. Proc. Natl. Acad. Sci. USA 2012, 109, 15888–15893. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nogueiras, R.; Habegger, K.M.; Chaudhary, N.; Finan, B.; Banks, A.S.; Dietrich, M.O.; Horvath, T.L.; Sinclair, D.A.; Pfluger, P.T.; Tschöp, M.H. Sirtuin 1 and sirtuin 3: Physiological modulators of metabolism. Physiol. Rev. 2012, 92, 1479–1514. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Mulcahy Levy, J.M.; Thorburn, A. Autophagy in cancer: Moving from understanding mechanism to improving therapy responses in patients. Cell Death Differ. 2020, 27, 843–857. [Google Scholar] [CrossRef] [PubMed]
- White, E. The role for autophagy in cancer. J. Clin. Investig. 2015, 125, 42–46. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Han, W.; Yu, F.; Cao, J.; Dong, B.; Guan, W.; Shi, J. Valproic Acid Enhanced Apoptosis by Promoting Autophagy Via Akt/mTOR Signaling in Glioma. Cell Transpl. 2020, 29, 1–10. [Google Scholar] [CrossRef]
- Wang, X.; Hu, S.; Liu, L. Phosphorylation and acetylation modifications of FOXO3a: Independently or synergistically? Oncol. Lett. 2017, 13, 2867–2872. [Google Scholar] [CrossRef] [Green Version]
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Jang, E.-H.; Lee, J.-H.; Kim, S.-A. Acute Valproate Exposure Induces Mitochondrial Biogenesis and Autophagy with FOXO3a Modulation in SH-SY5Y Cells. Cells 2021, 10, 2522. https://doi.org/10.3390/cells10102522
Jang E-H, Lee J-H, Kim S-A. Acute Valproate Exposure Induces Mitochondrial Biogenesis and Autophagy with FOXO3a Modulation in SH-SY5Y Cells. Cells. 2021; 10(10):2522. https://doi.org/10.3390/cells10102522
Chicago/Turabian StyleJang, Eun-Hye, Jung-Ho Lee, and Soon-Ae Kim. 2021. "Acute Valproate Exposure Induces Mitochondrial Biogenesis and Autophagy with FOXO3a Modulation in SH-SY5Y Cells" Cells 10, no. 10: 2522. https://doi.org/10.3390/cells10102522