{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,18]],"date-time":"2026-04-18T02:37:26Z","timestamp":1776479846972,"version":"3.51.2"},"reference-count":48,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,10,4]],"date-time":"2022-10-04T00:00:00Z","timestamp":1664841600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Gastric cancer (GC) is one of the most malignant and lethal cancers worldwide. Multiple microRNAs (miRNAs) have been identified as key regulators in the progression of GC. However, the underlying pathogenesis that miRNAs govern GC malignancy remains uncertain. Here, we identified a novel miR-585-5p as a key regulator in GC development.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The expression of miR-585-5p in the context of GC tissue was detected by <jats:italic>in situ<\/jats:italic> hybridization for GC tissue microarray and assessed by H-scoring. The gain- and loss-of-function analyses comprised of Cell Counting Kit-8 assay and Transwell invasion and migration assay. The expression of downstream microphthalmia-associated transcription factor (MITF), cyclic AMP-responsive element-binding protein 1 (CREB1) and mitogen-activated protein kinase 1 (MAPK1) were examined by Immunohistochemistry, quantitative real-time PCR and western blot. The direct regulation between miR-585-5p and MITF\/CREB1\/MAPK1 were predicted by bioinformatic analysis and screened by luciferase reporter assay. The direct transcriptional activation of CREB1 on MITF was verified by luciferase reporter assay, chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays (EMSAs). The interaction between MAPK1 and MITF was confirmed by co-immunoprecipitation (Co-IP) and immunofluorescent double-labelled staining.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>MiR-585-5p is progressively downregulated in GC tissues and low miR-585-5p levels were strongly associated with poor clinical outcomes. Further gain- and loss-of-function analyses showed that miR-585-5p possesses strong anti-proliferative and anti-metastatic capacities in GC. Follow-up studies indicated that miR-585-5p targets the downstream molecules CREB1 and MAPK1 to regulate the transcriptional and post-translational regulation of MITF, respectively, thus controlling its expression and cancer-promoting activity. MiR-585-5p directly and negatively regulates MITF together with CREB1 and MAPK1. According to bioinformatic analysis, promotor reporter gene assays, ChIP and EMSAs, CREB1 binds to the promotor region to enhance transcriptional expression of MITF. Co-IP and immunofluorescent double-labelled staining confirmed interaction between MAPK1 and MITF. Protein immunoprecipitation revealed that MAPK1 enhances MITF activity <jats:italic>via<\/jats:italic> phosphorylation (Ser73). MiR-585-5p can not only inhibit MITF expression directly, but also hinder MITF expression and pro-cancerous activity in a CREB1-\/MAPK1-dependent manner indirectly.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>In conclusion, this study uncovered miR-585-5p impedes gastric cancer proliferation and metastasis by orchestrating the interactions among CREB1, MAPK1 and MITF.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2022.1008195","type":"journal-article","created":{"date-parts":[[2022,10,4]],"date-time":"2022-10-04T04:51:01Z","timestamp":1664859061000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":7,"title":["MiR-585-5p impedes gastric cancer proliferation and metastasis by orchestrating the interactions among CREB1, MAPK1 and MITF"],"prefix":"10.3389","volume":"13","author":[{"given":"Yunwei","family":"Wang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiaoxia","family":"Zeng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yunshu","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zengshan","family":"Li","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sijun","family":"Hu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fangfang","family":"Yang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Na","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wenlan","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Tie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2022,10,4]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"394","DOI":"10.3322\/caac.21492","article-title":"Global cancer statistics 2018: Globocan estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"68","author":"Bray","year":"2018","journal-title":"CA Cancer J Clin"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1016\/s0140-6736(20)31288-5","article-title":"Gastric cancer","volume":"396","author":"Smyth","year":"2020","journal-title":"Lancet"},{"key":"B3","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1055\/s-0034-1397344","article-title":"Overview of microrna biology","volume":"35","author":"Mohr","year":"2015","journal-title":"Semin Liver Dis"},{"key":"B4","doi-asserted-by":"publisher","first-page":"e1000879","DOI":"10.1371\/journal.pgen.1000879","article-title":"Mir-218 inhibits invasion and metastasis of gastric cancer by targeting the Robo1 receptor","volume":"6","author":"Tie","year":"2010","journal-title":"PloS Genet"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.gene.2017.03.022","article-title":"Mir-218 inhibited tumor angiogenesis by targeting Robo1 in gastric cancer","volume":"615","author":"Zhang","year":"2017","journal-title":"Gene"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1210919","article-title":"Human papillomavirus type 16 reduces the expression of microrna-218 in cervical carcinoma cells","volume":"27","author":"Martinez","year":"2008","journal-title":"Oncogene"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkv703","article-title":"The microrna-183 cluster: The family that plays together stays together","volume":"43","author":"Dambal","year":"2015","journal-title":"Nucleic Acids Res"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.2147\/ott.s203175","article-title":"Psme3downregulated mir-585-3p promotes cell growth and proliferation in colon cancer by upregulating","volume":"12","author":"Liu","year":"2019","journal-title":"OncoTargets Ther"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1007\/s12094-016-1562-5","article-title":"Microrna-585 acts as a tumor suppressor in non-Small-Cell lung cancer by targeting hsmg-1","volume":"19","author":"Ding","year":"2017","journal-title":"Clin Trans Oncol"},{"key":"B10","doi-asserted-by":"publisher","first-page":"469","DOI":"10.1186\/s12935-020-01528-w","article-title":"Microrna-585 inhibits human glioma cell proliferation by directly targeting Mdm2","volume":"20","author":"Chen","year":"2020","journal-title":"Cancer Cell Int"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2018.03.116","article-title":"Microrna-585 suppresses tumor proliferation and migration in gastric cancer by directly targeting Mapk1","volume":"499","author":"Hu","year":"2018","journal-title":"Biochem Biophys Res Commun"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1111\/pcmr.12611","article-title":"Beyond mitf: Multiple transcription factors directly regulate the cellular phenotype in melanocytes and melanoma","volume":"30","author":"Seberg","year":"2017","journal-title":"Pigment Cell Melanoma Res"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1111\/his.12662","article-title":"Comparison of immunoreactive score, Her2\/Neu score and h score for the immunohistochemical evaluation of somatostatin receptors in bronchopulmonary neuroendocrine neoplasms","volume":"67","author":"Specht","year":"2015","journal-title":"Histopathology"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/nrg.2016.134","article-title":"A network-biology perspective of microrna function and dysfunction in cancer","volume":"17","author":"Bracken","year":"2016","journal-title":"Nat Rev Genet"},{"key":"B15","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1111\/j.1600-0749.2006.00348.x","article-title":"Activation of the mitf promoter by lipid-stimulated activation of P38-stress signalling to creb","volume":"19","author":"Saha","year":"2006","journal-title":"Pigment Cell Res"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1101\/gad.14.3.301","article-title":"C-kit triggers dual phosphorylations, which couple activation and degradation of the essential melanocyte factor mi","volume":"14","author":"Wu","year":"2000","journal-title":"Genes Dev"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-014-1791-0","article-title":"Mitf in melanoma: Mechanisms behind its expression and activity","volume":"72","author":"Hartman","year":"2015","journal-title":"Cell Mol Life Sciences: CMLS"},{"key":"B18","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/j.lfs.2016.08.015","article-title":"The activation of melanogenesis by p-creb and mitf signaling with extremely low-frequency electromagnetic fields on B16f10 melanoma","volume":"162","author":"Kim","year":"2016","journal-title":"Life Sci"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.7150\/thno.41502","article-title":"Jnk suppresses melanogenesis by interfering with creb-regulated transcription coactivator 3-dependent mitf expression","volume":"10","author":"Kim","year":"2020","journal-title":"Theranostics"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1111\/j.1755-148X.2010.00670.x","article-title":"Cell cycle inhibitor P21\/Waf1\/Cip1 as a cofactor of mitf expression in melanoma cells","volume":"23","author":"Sest\u00e1kov\u00e1","year":"2010","journal-title":"Pigment Cell Melanoma Res"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1038\/cdd.2008.148","article-title":"Tumour cell survival signalling by the Erk1\/2 pathway","volume":"16","author":"Balmanno","year":"2009","journal-title":"Cell Death Differentiation"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2021.641981","article-title":"De-ubiquitinating enzymes Usp21 regulate Mapk1 expression by binding to transcription factor Gata3 to regulate tumor growth and cell stemness of gastric cancer","volume":"9","author":"Guo","year":"2021","journal-title":"Front Cell Dev Biol"},{"key":"B23","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1186\/s12943-021-01358-y","article-title":"A novel protein encoded by Circmapk1 inhibits progression of gastric cancer by suppressing activation of mapk signaling","volume":"20","author":"Jiang","year":"2021","journal-title":"Mol Cancer"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.2147\/ott.s268554","article-title":"\u03b1-Msh-Pe38kdel kills melanoma cells Via modulating Erk1\/2\/Mitf\/Tyr signaling in an Mc1r-dependent manner","volume":"13","author":"Liu","year":"2020","journal-title":"OncoTargets Ther"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1016\/j.gene.2020.144937","article-title":"Micrornas in gastric cancer: Biomarkers and therapeutic targets","volume":"757","author":"Ghafouri-Fard","year":"2020","journal-title":"Gene"},{"key":"B26","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1186\/s12943-020-01185-7","article-title":"Noncoding rnas in gastric cancer: Implications for drug resistance","volume":"19","author":"Wei","year":"2020","journal-title":"Mol Cancer"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1007\/s10238-021-00704-0","article-title":"Research on correlations of mir-585 expression with progression and prognosis of triple-negative breast cancer","volume":"22","author":"Yao","year":"2021","journal-title":"Clin Exp Med"},{"key":"B28","doi-asserted-by":"publisher","first-page":"22270","DOI":"10.1038\/s41598-020-79337-4","article-title":"Identification and validation of a mirna-based prognostic signature for cervical cancer through an integrated bioinformatics approach","volume":"10","author":"Qi","year":"2020","journal-title":"Sci Rep"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1002\/path.5467","article-title":"Exome sequencing identifies new somatic alterations and mutation patterns of tongue squamous cell carcinoma in a Chinese population","volume":"251","author":"Zhang","year":"2020","journal-title":"J Pathol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.2147\/cmar.s212180","article-title":"Circular rna Hsa_Circrna_102958 promotes tumorigenesis of colorectal cancer Via mir-585\/Cdc25b axis","volume":"11","author":"Li","year":"2019","journal-title":"Cancer Manage Res"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1002\/cbin.11287","article-title":"Linc01436, regulating mir-585 and Fbxo11, is an oncogenic lncrna in the progression of gastric cancer","volume":"44","author":"Zhang","year":"2020","journal-title":"Cell Biol Int"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.18632\/aging.102112","article-title":"Downregulation of microrna-218 is cardioprotective against cardiac fibrosis and cardiac function impairment in myocardial infarction by binding to mitf","volume":"11","author":"Qian","year":"2019","journal-title":"Aging"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.4161\/rna.28865","article-title":"Microrna-218 inhibits melanogenesis by directly suppressing microphthalmia-associated transcription factor expression","volume":"11","author":"Guo","year":"2014","journal-title":"RNA Biol"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1007\/s11033-014-3351-y","article-title":"Mir-218 contributes to the transformation of 5-Aza\/Gf induced umbilical cord mesenchymal stem cells into hematopoietic cells through the mitf pathway","volume":"41","author":"Hu","year":"2014","journal-title":"Mol Biol Rep"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.29107","article-title":"Cse1l silence inhibits the growth and metastasis in gastric cancer by repressing gpnmb Via positively regulating transcription factor mitf","volume":"235","author":"Li","year":"2020","journal-title":"J Cell Physiol"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1042\/bsr20190066","article-title":"Mir-876-5p regulates gastric cancer cell proliferation, apoptosis and migration through targeting Wnt5a and mitf","volume":"39","author":"Xu","year":"2019","journal-title":"Biosci Rep"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0625.2011.01296.x","article-title":"Manassantin a inhibits camp-induced melanin production by down-regulating the gene expressions of mitf and tyrosinase in melanocytes","volume":"20","author":"Lee","year":"2011","journal-title":"Exp Dermatol"},{"key":"B38","doi-asserted-by":"publisher","first-page":"983","DOI":"10.1101\/gad.324657.119","article-title":"Mitf-the first 25 years","volume":"33","author":"Goding","year":"2019","journal-title":"Genes Dev"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1038\/jid.2014.548","article-title":"P21-activated kinase 4 critically regulates melanogenesis Via activation of the Creb\/Mitf and beta;-Catenin\/Mitf pathways","volume":"135","author":"Yun","year":"2015","journal-title":"J Invest Dermatol"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1111\/exd.13836","article-title":"Mannosylerythritol lipids inhibit melanogenesis Via suppressing erk-Creb-Mitf-Tyrosinase signalling in normal human melanocytes and a three-dimensional human skin equivalent","volume":"28","author":"Bae","year":"2019","journal-title":"Exp Dermatol"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.3390\/ijms19020409","article-title":"Antioxidative and anti-melanogenic activities of bamboo stems (Phyllostachys nigra variety henosis) Via Pka\/Creb-mediated mitf downregulation in B16f10 melanoma cells","volume":"19","author":"Choi","year":"2018","journal-title":"Int J Mol Sci"},{"key":"B42","doi-asserted-by":"publisher","first-page":"298","DOI":"10.1038\/34681","article-title":"Map kinase links the transcription factor microphthalmia to c-kit signalling in melanocytes","volume":"391","author":"Hemesath","year":"1998","journal-title":"Nature"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1810498115","article-title":"Braf\/Mapk and Gsk3 signaling converges to control mitf nuclear export","volume":"115","author":"Ngeow","year":"2018","journal-title":"Proc Natl Acad Sci U.S.A."},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2008.05.022","article-title":"Ubc9 sumoylation regulates sumo target discrimination","volume":"31","author":"Knipscheer","year":"2008","journal-title":"Mol Cell"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2018.05.083","article-title":"Sargaquinoic acid ameliorates hyperpigmentation through camp and erk-mediated downregulation of mitf in \u03b1-Msh-Stimulated B16f10 cells","volume":"104","author":"Azam","year":"2018","journal-title":"Biomed Pharmacother = Biomed Pharmacotherapie"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200505059","article-title":"Elevated expression of mitf counteracts B-Raf\u2013stimulated melanocyte and melanoma cell proliferation","volume":"170","author":"Wellbrock","year":"2005","journal-title":"J Cell Biol"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.3390\/cancers12113166","article-title":"Targeting creb in cancer therapy: A key candidate or one of many","volume":"12","author":"Sapio","year":"2020","journal-title":"Update Cancers"},{"key":"B48","doi-asserted-by":"publisher","first-page":"1999","DOI":"10.1007\/s00125-017-4356-6","article-title":"Erk1 is dispensable for mouse pancreatic beta cell function but is necessary for glucose-induced full activation of Msk1 and creb","volume":"60","author":"Leduc","year":"2017","journal-title":"Diabetologia"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.1008195\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,10,4]],"date-time":"2022-10-04T04:51:05Z","timestamp":1664859065000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.1008195\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,4]]},"references-count":48,"alternative-id":["10.3389\/fimmu.2022.1008195"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2022.1008195","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,4]]},"article-number":"1008195"}}