{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T18:55:29Z","timestamp":1783104929317,"version":"3.54.6"},"reference-count":103,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2024,2,15]],"date-time":"2024-02-15T00:00:00Z","timestamp":1707955200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006595","name":"Ministry of Research, Innovation and Digitization, CCCDI-UEFISCDI","doi-asserted-by":"publisher","award":["PN-III-P4-PCE-2021-0549"],"award-info":[{"award-number":["PN-III-P4-PCE-2021-0549"]}],"id":[{"id":"10.13039\/501100006595","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006595","name":"Ministry of Research, Innovation and Digitization, CCCDI-UEFISCDI","doi-asserted-by":"publisher","award":["PN 23.16.01.03"],"award-info":[{"award-number":["PN 23.16.01.03"]}],"id":[{"id":"10.13039\/501100006595","id-type":"DOI","asserted-by":"publisher"}]},{"name":"PNCDI III","award":["PN-III-P4-PCE-2021-0549"],"award-info":[{"award-number":["PN-III-P4-PCE-2021-0549"]}]},{"name":"PNCDI III","award":["PN 23.16.01.03"],"award-info":[{"award-number":["PN 23.16.01.03"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In the present study, we employed the ddPCR and IHC techniques to assess the prevalence and roles of RAS and RAF mutations in a small batch of melanoma (n = 22), benign moles (n = 15), and normal skin samples (n = 15). Mutational screening revealed the coexistence of BRAF and NRAS mutations in melanomas and nevi and the occurrence of NRAS G12\/G13 variants in healthy skin. All investigated nevi had driver mutations in the BRAF or NRAS genes and elevated p16 protein expression, indicating cell cycle arrest despite an increased mutational burden. BRAF V600 mutations were identified in 54% of melanomas, and NRAS G12\/G13 mutations in 50%. The BRAF mutations were associated with the Breslow index (BI) (p = 0.029) and TIL infiltration (p = 0.027), whereas the NRAS mutations correlated with the BI (p = 0.01) and the mitotic index (p = 0.04). Here, we demonstrate that the \u201cyoung\u201d ddPCR technology is as effective as a CE-IVD marked real-time PCR method for detecting BRAF V600 hotspot mutations in tumor biopsies and recommend it for extended use in clinical settings. Moreover, ddPCR was able to detect low-frequency hotspot mutations, such as NRAS G12\/G13, in our tissue specimens, which makes it a promising tool for investigating the mutational landscape of sun-damaged skin, benign nevi, and melanomas in more extensive clinical studies.<\/jats:p>","DOI":"10.3390\/ijms25042308","type":"journal-article","created":{"date-parts":[[2024,2,15]],"date-time":"2024-02-15T05:55:59Z","timestamp":1707976559000},"page":"2308","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6464-381X","authenticated-orcid":false,"given":"Elena-Georgiana","family":"Dobre","sequence":"first","affiliation":[{"name":"Doctoral School, Faculty of Biology, University of Bucharest, 050095 Bucharest, Romania"},{"name":"\u201cVictor Babes\u201d National Institute of Pathology, 050096 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7377-3325","authenticated-orcid":false,"given":"Luciana","family":"Nichita","sequence":"additional","affiliation":[{"name":"\u201cVictor Babes\u201d National Institute of Pathology, 050096 Bucharest, Romania"},{"name":"Colentina Clinical Hospital, 020125 Bucharest, Romania"},{"name":"Department of Pathology, Faculty of Dental Medicine, \u201cCarol Davila\u201d University of Medicine and Pharmacy, 020021 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristiana","family":"Popp","sequence":"additional","affiliation":[{"name":"\u201cVictor Babes\u201d National Institute of Pathology, 050096 Bucharest, Romania"},{"name":"Colentina Clinical Hospital, 020125 Bucharest, Romania"},{"name":"Department of Pathology, Faculty of Dental Medicine, \u201cCarol Davila\u201d University of Medicine and Pharmacy, 020021 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3498-8454","authenticated-orcid":false,"given":"Sabina","family":"Zurac","sequence":"additional","affiliation":[{"name":"\u201cVictor Babes\u201d National Institute of Pathology, 050096 Bucharest, Romania"},{"name":"Colentina Clinical Hospital, 020125 Bucharest, Romania"},{"name":"Department of Pathology, Faculty of Dental Medicine, \u201cCarol Davila\u201d University of Medicine and Pharmacy, 020021 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9339-2805","authenticated-orcid":false,"given":"Monica","family":"Neagu","sequence":"additional","affiliation":[{"name":"Doctoral School, Faculty of Biology, University of Bucharest, 050095 Bucharest, Romania"},{"name":"\u201cVictor Babes\u201d National Institute of Pathology, 050096 Bucharest, Romania"},{"name":"Colentina Clinical Hospital, 020125 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.jdin.2020.10.013","article-title":"The global burden of skin cancer: A longitudinal analysis from the Global Burden of Disease Study, 1990\u20132017","volume":"2","author":"Urban","year":"2021","journal-title":"JAAD Int."},{"key":"ref_2","first-page":"591","article-title":"Vital signs: Melanoma incidence and mortality trends and projections\u2014United States, 1982\u20132030","volume":"64","author":"Guy","year":"2015","journal-title":"MMWR Morb. 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