MiR-130a Acts as a Tumor Suppressor MicroRNA in Cutaneous Squamous Cell Carcinoma and Regulates the Activity of the BMP/SMAD Pathway by Suppressing ACVR1

Cutaneous squamous cell carcinoma (cSCC) is a malignant neoplasm of the skin resulting from accumulation somatic mutations due to solar radiation. cSCC one fastest increasing malignancies, and it represents particular problem among immunosuppressed individuals. MicroRNAs are short noncoding RNAs that regulate expression protein-coding genes at post-transcriptional level. In this study, we identify miR-130a be downregulated in compared healthy precancerous lesions (actinic keratosis). Moreoever, show its regulated transcriptional level by HRAS MAPK signaling pathway. We demonstrate overexpession suppresses long-term capacity growth, motility invasion ability human lines. report growth xenografts mice. Mechanistically, directly targets ACVR1 (ALK2), changes levels result decreased activity BMP/SMAD pathway through ACVR1. These data reveal link between activated miR-130a, which acts as tumor-suppressor microRNA contribute better understanding molecular processes during transformation epidermal keratinocytes. most common malignancies affecting more than 500,000 new patients per year throughout world. The incidence fast Caucasian population (Alam Ratner, 2001Alam M. Ratner D. squamous-cell carcinoma.N Engl J Med. 2001; 344: 975-983Crossref PubMed Scopus (928) Google Scholar; Que et al., 2018Que S.K.T. Zwald F.O. Schmults C.D. carcinoma: incidence, risk factors, diagnosis, staging.J Am Acad Dermatol. 2018; 78: 237-247Abstract Full Text PDF (215) Scholar). primary factor for chronic exposure UVR, induces epigenetic alterations transformation. can arise actinic keratosis (AK), commonly seen on sun-damaged skin, diagnosed with AK have 10% lifetime develop cSCCs (Salasche, 2000Salasche S.J. 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Results qRT-PCR-analysis demonstrated was significantly both normal (Figure 1a) AK, suggesting associated phenotype. situ hybridization using locked nucleic acid probes samples evenly expressed all layers compartment 1b). contrast, undetectable samples, tumorigenesis. aggressive (Pickering 2014Pickering C.R. Zhou Lee J.J. Drummond J.A. Peng S.A. Saade R.E. al.Mutational landscape 20: 6582-6592Crossref (333) hypothesized modulation could affect miR-130a. test hypothesis, small interfering RNA?mediated knockdown UT-SCC-7 line, which, expected, protein phosphorylation extracellular signal–regulated kinase (ERK), downstream effector 1c). qRT-PCR depletion cells transient 1d). further HRAS/MAPK/ERK (MEK)/ERK1/2 expression, were treated U0126 (a selective inhibitor kinases, MEK1 MEK2 [Favata 1998Favata M.F. Horiuchi K.Y. Manos E.J. Daulerio A.J. Stradley D.A. 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Thus, tumor-Suppressor single grow critical characteristic cells, required establishment metastases. this, assay A431 S2a) colony-forming S2c d). Furthermore, sphere evaluate growth. size 2c Conversely, inhibition endogenous larger spheres 2e f Supplementary S2b). modulates regulating Next, investigated essential features disease. scratch wound-healing Measurement wound area inhibited closure healing assays 3a). result, migration 3b) 3c) transwell assays. contrast miR-130a-overexpression, comparison sequences 3d). Inhibition also greatly migratory 3e) 3f) suppresse invasiveness vitro. next sought determine basis effects regulates. interrogated recent RNA-sequencing (Das Mahapatra 2020Das Pasquali Søndergaard J.N. Lapins Nemeth I.B. Baltás al.A comprehensive coding non-coding transcriptomic carcinoma.Sci 2020; 10: 1-12Crossref (26) Scholar) miR-130a?target prediction miRNA-binding sites 3?UTRs TargetScan S3a). 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Consistent findings, immunofluorescence p-SMAD1 nucleus 4g). staining analyze whether modulate SMAD-dependent DNA-binding reporter pGL3-BRE-luc plasmid, BMP-responsive construct respond p-SMAD1, p-SMAD5, p-SMAD8 (Korchynskyi ten Dijke, 2002Korchynskyi Dijke distinct critically important bone morphogenetic protein-specific response elements Id1 promoter.J 2002; 277: 4883-4891Abstract (677) promoter-luciferase promoter 4h), thus corroborate conclusion mediate cell-migration carried after Knockdown western 5a), p-SMAD1/5 5a). similar 5b). recapitulating miR-130 overexpression, silencing reduced migrating Transwell inserts 5c). counteract miR-130a-overexpression. transfected ACVR1-plasmid containing open reading frame without (pCMV-ACVR1), refer miR-130a-resistant miR-130a–resistant S4a) SMAD1/5 S4b) S5a b). HRAS–MEK–ERK1/2. turn, pathway, 6). involved related vivo. consequence BMP/SMAD1 showing suggested Interestingly, recently differences (Lambert 2014Lambert Mladkova Gulati Hamoudi R. 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Eidsmo Nagy al.MiR-125b, psoriasis, keratinocyte proliferation FGFR2.J 131: 1521-1529Abstract (155) miRNeasy FFPE Kit (Qiagen, Sollentuna, Sweden), respectively. details culture, transfections transductions, qRT-PCR, hybridization, experiment, immunostaining, blot, assay, statistical available Materials Methods online. Data sharing applicable article no sets generated analyzed supporting Materials. Warangkana Lohcharoenkal: https://orcid.org/0000-0001-6541-1693 Li: https://orcid.org/0000-0002-7199-4298 Kunal Das Mahapatra: https://orcid.org/0000-0002-2831-0751 Jan Lapins: https://orcid.org/0000-0002-0882-8959 Bernhard Homey: https://orcid.org/0000-0002-5784-4146 Enik? Sonkoly: https://orcid.org/0000-0002-4909-5413 Andor Pivarcsi: https://orcid.org/0000-0003-2196-1102 authors state conflict interest. supported Swedish Skin Research Foundation (Hudfonden/Welander Finsens Foundation), Society (Cancerfonden, LEO (Ballerup, Denmark), Council (Vetenskapsrådet, Stockholm County (Sweden) (SLL), Avtal om Läkarutbildning och Forskning (ALF Medicin), Deutsche Forschungsgemeinschaft (DFG) (FOR2690 Ho 2092/7-1 ALF Medicine 20190367). Conceptualization: AP, ES, WL; Curation: WL, CL, KDM; Formal Analysis: KDM, AP; Funding Acquisition: ES; Investigation: Project Administration: Resources: JL, BH, Supervision: Validation: CL; Visualization: Writing - Original Draft Preparation: Review Editing: AP Download .pdf (.7 MB) Help pdf files