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Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR
oleh: Elena-Georgiana Dobre, Luciana Nichita, Cristiana Popp, Sabina Zurac, Monica Neagu
Format: | Article |
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Diterbitkan: | MDPI AG 2024-02-01 |
Deskripsi
In the present study, we employed the ddPCR and IHC techniques to assess the prevalence and roles of <i>RAS</i> and <i>RAF</i> mutations in a small batch of melanoma (<i>n</i> = 22), benign moles (<i>n</i> = 15), and normal skin samples (<i>n</i> = 15). Mutational screening revealed the coexistence of <i>BRAF</i> and <i>NRAS</i> mutations in melanomas and nevi and the occurrence of <i>NRAS</i> G12/G13 variants in healthy skin. All investigated nevi had driver mutations in the <i>BRAF</i> or <i>NRAS</i> genes and elevated p16 protein expression, indicating cell cycle arrest despite an increased mutational burden. <i>BRAF</i> V600 mutations were identified in 54% of melanomas, and <i>NRAS</i> G12/G13 mutations in 50%. The <i>BRAF</i> mutations were associated with the Breslow index (BI) (<i>p</i> = 0.029) and TIL infiltration (<i>p</i> = 0.027), whereas the <i>NRAS</i> mutations correlated with the BI (<i>p</i> = 0.01) and the mitotic index (<i>p</i> = 0.04). Here, we demonstrate that the “young” ddPCR technology is as effective as a CE-IVD marked real-time PCR method for detecting <i>BRAF</i> 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 <i>NRAS</i> 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.