Expression of Molecular Markers in Aspirates of Patients with BRAF-negative Cutaneous Melanoma: A Case–Control Study



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Abstract

BACKGROUND: Cutaneous melanoma is one of the most common malignant tumors of the skin.

AIM: To investigate the clinical and morphological features, as well as the expression of transcription factors, growth factors, and components of the AKT/m-TOR signaling pathway in aspirates obtained from tumor tissue following fine-needle biopsy in patients with cutaneous and mucosal melanoma.

METHODS: The study included 41 patients with verified melanoma of the skin at various sites and mucosal melanoma of the nasal cavity, stages T1a–4bN0M0 (I–IV), as well as 18 patients with cutaneous nevi at different sites. The age of the patients ranged from 45 to 72 years; 25 patients were men (62%) and 16 were women (38%). Expression of signaling cascade components was evaluated using real-time polymerase chain reaction. The BRAF mutation status was determined using allele-specific real-time polymerase chain reaction. Statistical analysis was performed using nonparametric methods (Mann–Whitney U test; Kruskal–Wallis test). A p-value of <0.05 was considered statistically significant.

RESULTS: Aspirates from melanoma tissue showed increased expression of the following markers: AKT, 13.47-fold; c-RAF, 16.86-fold; m-TOR, 90.25-fold; PDK1, 63.2-fold; VEGFR2, 7.28-fold; CAIX, 801.69-fold; VHL, 398-fold; PD-L1, 28.18-fold; AMPK, 67.36-fold; and LC3B, 97-fold. In ulcerated tumors, a decrease in several important molecular markers was observed: 4EBP1, 13.39-fold (p = 0.0048); NF-kB p50, 19.38-fold (p = 0.0015); and VHL, 6.15-fold (p = 0.004). When comparing molecular marker expression by Clark’s level of invasion (from the epidermis to subcutaneous adipose tissue), AKT expression increased from 5.94-fold in group 3 to 56.96-fold in group 5, compared to group 2 (p = 0.0387). In group 3, there was also a marked increase in GSK-3β expression (30.49-fold) and PD-1 expression (90.8-fold; p = 0.0216), accompanied by a significant decrease in HIF-1 expression (891.44-fold; p = 0.004).

CONCLUSION: Aspirates from BRAF-negative tumors revealed evidence of autophagy activation and enhanced tumor immunogenicity. In particular, elevated AKT kinase levels were accompanied by increased expression of autophagosome protein and the PD-1 receptor.

About the authors

Veronika A. Bogdanova

Cancer Research Institute, Tomsk National Research Medical Center

Email: Vnika6906@yandex.ru
ORCID iD: 0009-0003-8473-4182
SPIN-code: 8903-4911

Assistant, Depart. of Biochemistry and Molecular Biology with the Course of KLD

Tomsk

Lyudmila V. Spirina

Siberian State Medical University; Cancer Research Institute, Tomsk National Research Medical Center

Email: spirinalvl@mail.ru
ORCID iD: 0000-0002-5269-736X
SPIN-code: 1336-8363

MD, Dr. Sci. (Medicine), Leading research associate, Tumor Biochemistry Lab., Professor, Depart. of Biochemistry and Molecular Biology with a Course in Clinical Laboratory Diagnostics

Russian Federation, Tomsk; Tomsk

Svetlana Yu. Chizhevskaya

Siberian State Medical University; Cancer Research Institute, Tomsk National Research Medical Center

Email: sch@oncology.tomsk.ru
ORCID iD: 0000-0003-2974-4778
SPIN-code: 9561-3382

MD, Dr. Sci. (Medicine), Leading research associate, Depart. of Head and Neck Tumors, Oncologist of the highest category

Russian Federation, Tomsk; Tomsk

Konstantin V. Nikulnikov

Siberian State Medical University

Author for correspondence.
Email: kast10sha91@mail.ru
ORCID iD: 0009-0004-7211-7686
SPIN-code: 5258-7989

oncologist, Depart. of Head and Neck Tumors, Research Institute of Oncology

Russian Federation, Tomsk

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