{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T14:39:58Z","timestamp":1787063998490,"version":"build-2736575974"},"reference-count":41,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,1,28]],"date-time":"2022-01-28T00:00:00Z","timestamp":1643328000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81801488"],"award-info":[{"award-number":["81801488"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Shanghai \u201cRising Stars of Medical Talent\u201d Youth Development Program-Youth Medical Tal-ents-Clinical Pharmacist Program","award":["SHWRS (2020)_087"],"award-info":[{"award-number":["SHWRS (2020)_087"]}]},{"DOI":"10.13039\/100017950","name":"Shanghai Municipal Health Commission","doi-asserted-by":"publisher","award":["20204Y0339"],"award-info":[{"award-number":["20204Y0339"]}],"id":[{"id":"10.13039\/100017950","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Insulin-like growth factor 1 (IGF-1) not only regulates neuronal function and development but also is neuroprotective in the setting of acute ischemic stroke. G-protein-coupled receptor 17 (GPR17) expression in brain tissue serves as an indicator of brain damage. As whether IGF-1 regulates GPR17 expression remains unknown, the aim of this study is to investigate how IGF-1 regulates GPR17 expression in vitro. Human neuroblastoma SK-N-SH cells were used. Lentivirus-mediated short hairpin RNA (shRNA) was constructed to mediate the silencing of FoxO1, while adenoviral vectors were used for its overexpression. Verification of the relevant signaling cascade was performed using a FoxO1 inhibitor (AS1842856), a phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002), and a GPR17 antagonist (cangrelor). Cell proliferation was analyzed using EdU staining; immunofluorescence staining was used to detect the expression and subcellular localization of FoxO1. Chromatin immunoprecipitation was used to analyze the binding of FoxO1 to the GPR17 promoter in SK-N-SH cells. The expression of FoxO1, GPR17, and protein kinase B (also known as Akt) mRNA and protein as well as the levels of FoxO1 and Akt phosphorylation were investigated in this study. IGF-1 was found to downregulate FoxO1 and GPR17 expression in SK-N-SH cells while promoting cell viability and proliferation. Inhibition of FoxO1 and antagonism of GPR17 were found to play a role similar to that of IGF-1. Silencing of FoxO1 by lentivirus-mediated shRNA resulted in the downregulation of FoxO1 and GPR17 expression. The overexpression of FoxO1 via adenoviral vectors resulted in the upregulation of FoxO1 and GPR17 expression. Blocking of PI3K signaling by LY294002 inhibited the effect of IGF-1 on GPR17 suppression. Results from chromatin immunoprecipitation revealed that IGF-1 promotes FoxO1 nuclear export and reduces FoxO1 binding to the GPR17 promoter in SK-N-SH cells. Here, we conclude that IGF-1 enhances cell viability and proliferation in SK-N-SH cells via the promotion of FoxO1 nuclear export and reduction of FoxO1 binding to the GPR17 promoter via PI3K\/Akt signaling. Our findings suggest that the enhancement of IGF-1 signaling to antagonize GPR17 serves as a potential therapeutic strategy in the management of acute ischemic stroke.<\/jats:p>","DOI":"10.3390\/ijms23031513","type":"journal-article","created":{"date-parts":[[2022,1,29]],"date-time":"2022-01-29T05:26:17Z","timestamp":1643433977000},"page":"1513","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Insulin-like Growth Factor 1 Promotes Cell Proliferation by Downregulation of G-Protein-Coupled Receptor 17 Expression via PI3K\/Akt\/FoxO1 Signaling in SK-N-SH Cells"],"prefix":"10.3390","volume":"23","author":[{"given":"Ka-Na","family":"Lin","sequence":"first","affiliation":[{"name":"Center for Brain Science & Clinical Research Center, Shanghai Children\u2019s Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Anesthesiology, Shanghai Children\u2019s Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Shanghai Children\u2019s Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shi-Ying","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Shanghai Children\u2019s Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Li","sequence":"additional","affiliation":[{"name":"Center for Brain Science & Clinical Research Center, Shanghai Children\u2019s Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China"},{"name":"Department of Pharmacy, Shanghai Children\u2019s Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1088","DOI":"10.1001\/jama.2020.26867","article-title":"Diagnosis and Management of Transient Ischemic Attack and Acute Ischemic Stroke: A Review","volume":"325","author":"Mendelson","year":"2021","journal-title":"JAMA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1056\/NEJMcp1917030","article-title":"Acute Ischemic Stroke","volume":"383","author":"Powers","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"l6983","DOI":"10.1136\/bmj.l6983","article-title":"Management of acute ischemic stroke","volume":"368","author":"Phipps","year":"2020","journal-title":"BMJ"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"100821","DOI":"10.1016\/j.yfrne.2020.100821","article-title":"Insulin-like growth factor 1 signaling in motor neuron and polyglutamine diseases: From molecular pathogenesis to therapeutic perspectives","volume":"57","author":"Pennuto","year":"2020","journal-title":"Front. Neuroendocrinol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1186\/s12929-019-0587-9","article-title":"The functional roles of IGF-1 variants in the susceptibility and clinical outcomes of mild traumatic brain injury","volume":"26","author":"Wang","year":"2019","journal-title":"J. Biomed. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"736104","DOI":"10.1155\/2015\/736104","article-title":"Role and Importance of IGF-1 in Traumatic Brain Injuries","volume":"2015","author":"Mangiola","year":"2015","journal-title":"Biomed Res. Int."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2260","DOI":"10.1161\/01.CIR.0000144309.87183.FB","article-title":"Insulin-like growth factor-1 as a vascular protective factor","volume":"110","author":"Mangiola","year":"2004","journal-title":"Circulation"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1126\/science.275.5300.661","article-title":"Regulation of neuronal survival by the serine-threonine protein kinase Akt","volume":"275","author":"Dudek","year":"1997","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"48","DOI":"10.3389\/fnana.2015.00048","article-title":"Neuroprotective effects of DAHP and Triptolide in focal cerebral ischemia via apoptosis inhibition and PI3K\/Akt\/mTOR pathway activation","volume":"9","author":"Li","year":"2015","journal-title":"Front. Neuroanat."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1016\/S0092-8674(00)80595-4","article-title":"Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor","volume":"96","author":"Brunet","year":"1999","journal-title":"Cell"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/S0736-5748(02)00047-3","article-title":"Functions of PI 3-kinase in development of the nervous system","volume":"20","author":"Rodgers","year":"2002","journal-title":"Int. J. Dev. Neurosci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/j.brainres.2018.02.049","article-title":"Intra-arterial human urinary kallidinogenase alleviates brain injury in rats with permanent middle cerebral artery occlusion through PI3K\/AKT\/FoxO1 signaling pathway","volume":"1687","author":"Ma","year":"2018","journal-title":"Brain Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4615","DOI":"10.1038\/sj.emboj.7601341","article-title":"The orphan receptor GPR17 identified as a new dual uracil nucleotides\/cysteinyl-leukotrienes receptor","volume":"25","author":"Ciana","year":"2006","journal-title":"EMBO J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1007\/s11302-013-9366-3","article-title":"Changes of the GPR17 receptor, a new target for neurorepair, in neurons and glial cells in patients with traumatic brain injury","volume":"9","author":"Franke","year":"2013","journal-title":"Purinergic Signal."},{"key":"ref_15","first-page":"2750","article-title":"GPR17 mediates ischemia-like neuronal injury via microglial activation","volume":"42","author":"Zhao","year":"2018","journal-title":"Int. J. Mol. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.neuroscience.2011.11.066","article-title":"The new P2Y-like receptor G protein-coupled receptor 17 mediates acute neuronal injury and late microgliosis after focal cerebral ischemia in rats","volume":"202","author":"Zhao","year":"2012","journal-title":"Neuroscience"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.lfs.2019.03.045","article-title":"The role of orphan G protein-coupled receptors in the pathophysiology of multiple sclerosis: A review","volume":"224","author":"Alavi","year":"2019","journal-title":"Life Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.neuropharm.2015.10.005","article-title":"CNS remyelination as a novel reparative approach to neurodegenerative diseases: The roles of purinergic signaling and the P2Y-like receptor GPR17","volume":"104","author":"Fumagalli","year":"2016","journal-title":"Neuropharmacology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2206","DOI":"10.1093\/brain\/awp147","article-title":"The P2Y-like receptor GPR17 as a sensor of damage and a new potential target in spinal cord injury","volume":"132","author":"Ceruti","year":"2009","journal-title":"Brain A J. Neurol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1314","DOI":"10.1016\/j.cell.2012.04.032","article-title":"FoxO1 target Gpr17 activates AgRP neurons to regulate food intake","volume":"149","author":"Ren","year":"2012","journal-title":"Cell"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1845","DOI":"10.1073\/pnas.1424968112","article-title":"GPR17 gene disruption does not alter food intake or glucose homeostasis in mice","volume":"112","author":"Mastaitis","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3670","DOI":"10.2337\/db15-0390","article-title":"Gpr17 in AgRP Neurons Regulates Feeding and Sensitivity to Insulin and Leptin","volume":"64","author":"Ren","year":"2015","journal-title":"Diabetes"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1080\/13543776.2019.1568990","article-title":"GPR17 receptor modulators and their therapeutic implications: Review of recent patents","volume":"29","author":"Marucci","year":"2019","journal-title":"Expert Opin. Ther. Pat."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Bonfanti, E., Bonifacino, T., Raffaele, S., Milanese, M., Morgante, E., Bonanno, G., Abbracchio, M.P., and Fumagalli, M. (2020). Abnormal Upregulation of GPR17 Receptor Contributes to Oligodendrocyte Dysfunction in SOD1 G93A Mice. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21072395"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1957","DOI":"10.1002\/glia.23807","article-title":"Regulation and signaling of the GPR17 receptor in oligodendroglial cells","volume":"68","author":"Lecca","year":"2020","journal-title":"Glia"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.1093\/ptj\/pzz125","article-title":"Combination Therapy with Hyperbaric Oxygen and Erythropoietin Inhibits Neuronal Apoptosis and Improves Recovery in Rats with Spinal Cord Injury","volume":"99","author":"Zhou","year":"2019","journal-title":"Phys. Ther."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.lfs.2016.02.035","article-title":"Neuroprotective effects of phytosterols and flavonoids from Cirsium setidens and Aster scaber in human brain neuroblastoma SK-N-SH cells","volume":"148","author":"Zhou","year":"2016","journal-title":"Life Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1111\/j.1600-079X.2008.00648.x","article-title":"Melatonin attenuates methamphetamine-induced deactivation of the mammalian target of rapamycin signaling to induce autophagy in SK-N-SH cells","volume":"46","author":"Kongsuphol","year":"2009","journal-title":"J. Pineal Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.brainres.2010.11.062","article-title":"Neuroprotective effects of KR-62980, a new PPAR\u03b3 agonist, against chemical ischemia-reperfusion in SK-N-SH cells","volume":"1372","author":"Kim","year":"2011","journal-title":"Brain Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.neubiorev.2016.01.001","article-title":"The therapeutic potential of insulin-like growth factor-1 in central nervous system disorders","volume":"63","author":"Costales","year":"2016","journal-title":"Neurosci. Biobehav. Rev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"365","DOI":"10.3389\/fnagi.2017.00365","article-title":"Insulin-Like Growth Factor-1 and Neuroinflammation","volume":"9","author":"Labandeira","year":"2017","journal-title":"Front. Aging Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6252","DOI":"10.3390\/ijms15046252","article-title":"A rapid and efficient immunoenzymatic assay to detect receptor protein interactions: G protein-coupled receptors","volume":"15","author":"Zappelli","year":"2014","journal-title":"Int. J. Mol. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Winter, N., Novatchkova, M., and Bachmair, A. (2021). Cellular Control of Protein Turnover via the Modification of the Amino Terminus. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073545"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1186\/s12974-021-02112-3","article-title":"Recombinant CCL17-dependent CCR4 activation alleviates neuroinflammation and neuronal apoptosis through the PI3K\/AKT\/Foxo1 signaling pathway after ICH in mice","volume":"18","author":"Deng","year":"2021","journal-title":"J. Neuroinflamm."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e269","DOI":"10.1002\/ctm2.269","article-title":"CD73 alleviates GSDMD-mediated microglia pyroptosis in spinal cord injury through PI3K\/AKT\/Foxo1 signaling","volume":"11","author":"Xu","year":"2021","journal-title":"Clin. Transl. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"7566365","DOI":"10.1155\/2021\/7566365","article-title":"CRNDE\/ETS1\/GPR17 Facilitates the Proliferation, Migration, and Invasion of Glioma","volume":"2021","author":"Hu","year":"2021","journal-title":"Comput. Math. Methods Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e364","DOI":"10.1161\/STR.0000000000000375","article-title":"2021 Guideline for the Prevention of Stroke in Patients With Stroke and Transient Ischemic Attack: A Guideline From the American Heart Association\/American Stroke Association","volume":"52","author":"Kleindorfer","year":"2021","journal-title":"Stroke"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1155","DOI":"10.1002\/glia.22506","article-title":"UDP-glucose enhances outward K(+) currents necessary for cell differentiation and stimulates cell migration by activating the GPR17 receptor in oligodendrocyte precursors","volume":"61","author":"Coppi","year":"2013","journal-title":"Glia"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1007\/s11302-016-9530-7","article-title":"A new role for the P2Y-like GPR17 receptor in the modulation of multipotency of oligodendrocyte precursor cells in vitro","volume":"12","author":"Boccazzi","year":"2016","journal-title":"Purinergic Signal."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1186\/s40001-021-00536-4","article-title":"Cangrelor ameliorates CLP-induced pulmonary injury in sepsis by inhibiting GPR17","volume":"26","author":"Luo","year":"2021","journal-title":"Eur. J. Med. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.intimp.2018.06.006","article-title":"Cangrelor alleviates pulmonary fibrosis by inhibiting GPR17-mediated inflammation in mice","volume":"62","author":"Zhan","year":"2018","journal-title":"Int. Immunopharmacol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1513\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:10:00Z","timestamp":1760134200000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1513"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,28]]},"references-count":41,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["ijms23031513"],"URL":"https:\/\/doi.org\/10.3390\/ijms23031513","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,28]]}}}