Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR

Dobre, Elena-Georgiana; Nichita, Luciana; Popp, Cristiana; Zurac, Sabina; Neagu, Monica

doi:10.3390/ijms25042308

Open AccessArticle

Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR

by

Elena-Georgiana Dobre

^1,2,†

,

Luciana Nichita

^2,3,4

,

Cristiana Popp

^2,3,4,

Sabina Zurac

^2,3,4

and

Monica Neagu

^1,2,3,*,†

¹

Doctoral School, Faculty of Biology, University of Bucharest, 050095 Bucharest, Romania

²

“Victor Babes” National Institute of Pathology, 050096 Bucharest, Romania

³

Colentina Clinical Hospital, 020125 Bucharest, Romania

⁴

Department of Pathology, Faculty of Dental Medicine, “Carol Davila” University of Medicine and Pharmacy, 020021 Bucharest, Romania

^*

Author to whom correspondence should be addressed.

^†

These authors contributed equally to this work.

Int. J. Mol. Sci. 2024, 25(4), 2308; https://doi.org/10.3390/ijms25042308

Submission received: 18 January 2024 / Revised: 10 February 2024 / Accepted: 13 February 2024 / Published: 15 February 2024

(This article belongs to the Section Molecular Pathology, Diagnostics, and Therapeutics)

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Abstract

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 “young” 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.

Keywords: BRAF V600E mutation; NRAS G12/G13 mutation; melanoma; nevi

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MDPI and ACS Style

Dobre, E.-G.; Nichita, L.; Popp, C.; Zurac, S.; Neagu, M. Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR. Int. J. Mol. Sci. 2024, 25, 2308. https://doi.org/10.3390/ijms25042308

AMA Style

Dobre E-G, Nichita L, Popp C, Zurac S, Neagu M. Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR. International Journal of Molecular Sciences. 2024; 25(4):2308. https://doi.org/10.3390/ijms25042308

Chicago/Turabian Style

Dobre, Elena-Georgiana, Luciana Nichita, Cristiana Popp, Sabina Zurac, and Monica Neagu. 2024. "Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR" International Journal of Molecular Sciences 25, no. 4: 2308. https://doi.org/10.3390/ijms25042308

APA Style

Dobre, E.-G., Nichita, L., Popp, C., Zurac, S., & Neagu, M. (2024). Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR. International Journal of Molecular Sciences, 25(4), 2308. https://doi.org/10.3390/ijms25042308

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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

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