Role of DARPP-32 in the Proliferation and Survival of Human Neural Stem Cells
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To investigate the role of dopamine- and cAMP-regulated neuronal phosphoprotein 32 kDa (DARPP-32), also known as protein phosphatase 1 regulatory subunit 1B (PPP1R1B), in cell proliferation and survival of human neural stem cells (hiNSCs) induced from human adipose-derived stem cells, DARPP-32 expression was knocked down by a lentiviral vector. Cell proliferation was relatively slower and doubling time was longer in the DARPP-32-knockdown hNSCs than in the scrambled control. Oxidative stress testing showed that DARPP-32 knockdown caused more cell death in hiNSCs after hydrogen peroxide (H2O2) treatment than in the scrambled control. The expressions of CTNNB1, CXCR4, and VCAM-1, which are related to cell proliferation, were lower in the DARPP-32-knockdown hiNSCs than in the scrambled control. In addition, DARPP-32 knockdown decreased the expression of Bcl-2 and increased the expression of Bax in response to H2O2 treatment. We also confirmed reduced expression of phosphorylated Akt (p-AKT) in DARPP-32-knockdown hiNSCs. In conclusion, these findings demonstrate the mechanisms by which DARPP-32 positively affects cell proliferation and survival in hiNSCs.
Abstract
To investigate the role of dopamine- and cAMP-regulated neuronal phosphoprotein 32 kDa (DARPP-32), also known as protein phosphatase 1 regulatory subunit 1B (PPP1R1B), in cell proliferation and survival of human neural stem cells (hiNSCs) induced from human adipose-derived stem cells, DARPP-32 expression was knocked down by a lentiviral vector. Cell proliferation was relatively slower and doubling time was longer in the DARPP-32-knockdown hNSCs than in the scrambled control. Oxidative stress testing showed that DARPP-32 knockdown caused more cell death in hiNSCs after hydrogen peroxide (H2O2) treatment than in the scrambled control. The expressions of CTNNB1, CXCR4, and VCAM-1, which are related to cell proliferation, were lower in the DARPP-32-knockdown hiNSCs than in the scrambled control. In addition, DARPP-32 knockdown decreased the expression of Bcl-2 and increased the expression of Bax in response to H2O2 treatment. We also confirmed reduced expression of phosphorylated Akt (p-AKT) in DARPP-32-knockdown hiNSCs. In conclusion, these findings demonstrate the mechanisms by which DARPP-32 positively affects cell proliferation and survival in hiNSCs.
참고문헌 (27)
- Toma JG, "Isolation of multipotent adult stem cells from the dermis of mammalian skin", Nat Cell Biol, 3, pp.778~784 (2001)[DOI]
- Oh JH, "Integrative analysis of genes and miRNA alterations in human embryonic stem cells-derived neural cells after exposure to silver nanoparticles", Toxicol Appl Pharmacol, 299, pp.8~23 (2016)[DOI]
- Kunii Y, "Revisiting DARPP-32in postmortem human brain: changes in schizophrenia and bipolar disorder and genetic associations with t-DARPP-32 expression", Mol Psychiatry, pp.192~199 (2014)[DOI]
- Reis HJ, "Is DARPP-32 a potential therapeutic target?", Expert Opin Ther Targets, 11(12), pp.1649~1661 (2007)[DOI]
- Lanet E, "Protection of Neuronal Diversity at the Expense of Neuronal Numbers during Nutrient Restriction in the Drosophila Visual System", Cell Rep, 3, pp.587~594 (2013)[DOI]
- Homem CF, "Proliferation control in neural stem and progenitor cells", Nat Rev Neurosci, 16(11), pp.647~659 (2015)[DOI]
- Park J, "Small molecule-based lineage switch of human adipose-derived stem cells into neural stem cells and functional GABAergic neurons", Sci Rep, 7(1), pp.10166~ (2017)[DOI]
- Yagishita S, "A critical time window for dopamine actions on the structural plasticity of dendritic spines", Science, 345(6204), pp.1616~1620 (2014)[DOI]
- Pfisterer U, "Neuronal survival in the brain: neuron type-specific mechanisms", Cell Death Dis., 8(3), pp.e2643~ (2017)[DOI]
- Green DR, "Means to an End: Apoptosis and other Cell Death Mechanisms" (2011), Cold Spring Harbor Laboratory Press
- Nakamura A, "Bcl-xL Is Essential for the Survival and Function of Differentiated Neurons in the Cortex That Control Complex Behaviors", J Neurosci., 36, pp.5448~5461 (2016)[DOI]
- Fang D, "Phosphorylation of β-catenin by AKT Promotes β-Catenin Transcriptional Activity", J Biol Chem., 282(15), pp.11221~11229 (2007)[DOI]
- Largaespada D, "Interweaving the Strands:β-Catenin, an HIV Co-Receptor, and Schwann Cell Tumors", Cancer Cell, 23(3), pp.269~271 (2013)[DOI]
- Badr HS, "Serum stromal-derived-factor-1 (CXCL12) and its alpha chemokine receptor (CXCR4) as biomarkers in neonatal sepsis", J Matern Fetal Neonatal Med., 31(16), pp.2209~2215 (2018)[DOI]
- Bagri A, "The chemokine SDF1regulates migration of dentate granule cells", Development, 129(18), pp.4249~4260 (2002)
- Zhu S, "Regulation of CXCR4-mediated invasion by DARPP-32 in gastric cancer cells", Mol Cancer Res., 11(1), pp.86~94 (2013)[DOI]
- Gao J, "Wnt", β-Catenin Signaling in Neural Stem Cell Homeostasis and Neurological Diseases, 2020, 1~15
- MacDonald BT, "Wnt", β-Catenin Signaling: Components, Mechanisms, and Diseases, 2009, pp.17(1), 9~26
- Pei Y, "WNT signaling increases proliferation and impairs differentiation of stem cells in the developing cerebellum", Development, 139, pp.1724~1733 (2012)[DOI]
- Vangamudi B, "Regulation of β-catenin by t-DARPP in upper gastrointestinal cancer cells", Mol Cancer, 10, pp.32~ (2011)[DOI]
- Zhao J, "A protocol for the culture and isolation of murine synovial fibroblasts", Biomed Rep., 5(2), pp.171~175 (2016)[DOI]
- Zhang J, "Knockdown of vascular cell adhesion molecule 1 impedes transforming growth factor beta 1-mediated proliferation, migration, and invasion of endometriotic cyst stromal cells", Reprod Biol Endocrinol, 17(1), pp.69~ (2019)[DOI]
- Alam SK, "DARPP-32 and t-DARPP promote non-small cell lung cancer growth through regulation of IKKα-dependent cell migration", Commun Biol., 1, pp.43~ (2018)[DOI]
- Belkhiri A, "t-Darpp promotes cancer cell survival by up-regulation of Bcl2 through Akt-dependent mechanism", Cancer Res., 68, pp.395~403 (2008)[DOI]
- Bianchi ME, "The Chemokine Receptor CXCR4 in Cell Proliferation and Tissue Regeneration", Front Immunol, 11, pp.2109~ (2020)[DOI]
- Delft MF, "How the Bcl-2 family of proteins interact to regulate apoptosis", Cell Research, 16(2), pp.203~213 (2006)[DOI]
- Dudek H, "Regulation of neuronal survival by the serine-threonine protein kinase Akt", Science, 275, pp.661~665 (1997)[DOI]