Analysis of MicroRNA Expression in Specimens From Patients With Cutaneous Melanoma

Importance: Cutaneous melanoma is regarded as the most lethal form of skin cancer. According to the literature, issues associated with accurate prognosis and early detection of relapses hinder effective treatment of melanomas. Over the past 20 years, microRNAs have been shown to modulate various biological processes. Disruption of microRNA expression and activity can contribute to the risk of cancer development, including cutaneous melanoma. Therefore, we decided to study some microRNAs as biomarkers for future use in cutaneous melanoma diagnosis.

Objective: To analyze the expression of microRNA-205, microRNA-199a-3p, microRNA-146a-5p in tumor tissue of patients diagnosed with cutaneous melanoma.

Materials and methods: We quantified the expression of microRNA-205, microRNA-199a-3p, microRNA-146a-5p using the realtime polymerase chain reaction in biopsy specimens from patients with melanoma (n=14). For all the specimens, the cycle threshold (Ct) value was 0.1 (qPCRsoft 3.0, Analytik Jena, Germany). Statistical analysis was performed using Statistica 7.0 (StatSoft, USA). Furthermore, we conducted a bioinformatics search for microRNA target genes using DIANA-mirPath v.3.0 (DIANA-Lab, Greece) and constructed gene interaction networks using STRING (https://string-db.org/).

Results: We detected 2 of 3 microRNAs in the group of patients with cutaneous melanoma: microRNA-205 and microRNA-146a-5p. In both the control group and the group of patients with melanoma, 1 of 3 microRNAs, namely miR-146a-5p, was found. The mean ∆Ct value of microRNA-146a-5p was 3.81±2.03 in the control group and 2.26±1.34 in the patients with melanoma. Due to the wide range of values in the control group and the group of patients with cutaneous melanoma, microRNA-146a-5p cannot be considered an informative biomarker. At the same time, we found high expression of microRNA-205 in the group of patients with cutaneous melanoma.

Conclusions: microRNA-205 was found to be valuable in melanoma diagnosis. However, further studies with larger sample size and additional stratification by histopathological features are needed.

1. Stefanova YuYu, Porkhanova NV, Murashko RA, Timoshkina NV, Maksimov AYu, Timofeeva SV. Mutational status of BRAF, NRAS genes in tumor material of patients diagnosed with cutaneous melanoma. Medical Genetics. 2024;23(7):51–56. (In Russ.).

2. NegriniM, FerracinM, SabbioniS, CroceCM. MicroRNAs in human cancer: from research to therapy. J Cell Sci. 2007;120(Pt 11):1833– 1840. PMID: 17515481. https://doi.org/10.1242/jcs.03450

3. Bertoli G, Cava C, Castiglioni I. MicroRNAs: new biomarkers for diagnosis, prognosis, therapy prediction and therapeutic tools for breast cancer. Theranostics. 2015;5(10):1122–1143. PMID: 26199650. PMCID: PMC4508501. https://doi.org/10.7150/thno.11543

4. Durante G, Broseghini E, Comito F, et al. Circulating microRNA biomarkers in melanoma and non-melanoma skin cancer. Expert Rev Mol Diagn. 2022;22(3):305–318. PMID: 35235479. https://doi.org/10.1080/14737159.2022.2049243

5. Petkevich AA, Shubina IZh, Abramov AA, Mamedova LT, Samoilenko IV, Kiselevsky MV. MicroRNA expression in melanocytes and melanoma cells. Russian Journal of Biotherapy. 2018;17(3):6–11. (In Russ.). https://doi.org/10.17650/1726-9784-2018-17-3-6-11

6. Dar AA, Majid S, de Semir D, Nosrati M, Bezrookove V, Kashani-Sabet M. miRNA-205 suppresses melanoma cell proliferation and induces senescence via regulation of E2F1 protein. J Biol Chem. 2011;286(19):16606–16614. PMID: 21454583. PMCID: PMC3089503. https://doi.org/10.1074/jbc.M111.227611

7. Liu S, Tetzlaff MT, Liu A, Liegl-Atzwanger B, Guo J, Xu X. Loss of microRNA-205 expression is associated with melanoma progression. Lab Invest. 2012;92(7):1084–1096. PMID: 22525428. https://doi.org/10.1038/labinvest.2012.62

8. Zhang Y, Zhao J, Ding M, et al. Loss of exosomal miR146a-5p from cancer-associated fibroblasts after androgen deprivation therapy contributes to prostate cancer metastasis. J Exp Clin Cancer Res. 2020;39(1):282. PMID: 33317606. PMCID: PMC7734763. https://doi.org/10.1186/s13046-020-01761-1

9. Ghafouri-Fard S, Gholipour M, Taheri M. MicroRNA Signature in melanoma: biomarkers and therapeutic targets. Front Oncol. 2021;11:608987. PMID: 33968718. PMCID: PMC8100681. https://doi.org/10.3389/fonc.2021.608987

10. Szklarczyk D, Kirsch R, Koutrouli M, et al. The STRING database in 2023: protein-protein association networks and functional enrichment analyses for any sequenced genome of interest. Nucleic Acids Res. 2023;51(D1):D638–D646. PMID: 36370105. PMCID: PMC9825434. https://doi.org/10.1093/nar/gkac1000

11. Hwang SJ, Seol HJ, Park YM, et al. MicroRNA-146a suppresses metastatic activity in brain metastasis. Mol Cells. 2012;34(3):329–334. PMID: 22949171. PMCID: PMC3887840. https://doi.org/10.1007/s10059-012-0171-6

12. Pu W, Shang Y, Shao Q, Yuan X. miR-146a promotes cell migration and invasion in melanoma by directly targeting SMAD4. Oncol Lett. 2018;15(5):7111–7117. PMID: 29731876. PMCID: PMC5921230. https://doi.org/10.3892/ol.2018.8172

13. Vergani E, Dugo M, Cossa M, et al. miR-146a-5p impairs melanoma resistance to kinase inhibitors by targeting COX2 and regulating NFkB-mediated inflammatory mediators. Cell Commun Signal. 2020;18(1):156. PMID: 32967672. PMCID: PMC7510138. https://doi.org/10.1186/s12964-020-00601-1

14. Pencheva N, Tran H, Buss C, et al. Convergent multimiRNA targeting of ApoE drives LRP1/LRP8-dependent melanoma metastasis and angiogenesis. Cell. 2012;151(5):1068–1082. PMID: 23142051. PMCID: PMC3753115. https://doi.org/10.1016/j.cell.2012.10.028

15. Sánchez-Sendra B, Serna E, Navarro L, et al. Transcriptomic identification of miR-205 target genes potentially involved in metastasis and survival of cutaneous malignant melanoma. Sci Rep. 2020;10(1):4771. PMID: 32179834. PMCID: PMC7075905. https://doi.org/10.1038/s41598-020-61637-4

16. Aksenenko M, Palkina N, Komina A, Tashireva L, Ruksha T. Differences in microRNA expression between melanoma and healthy adjacent skin. BMC Dermatol. 2019;5;19(1):1. PMID: 30611259. PMCID: PMC6321655. https://doi.org/10.1186/s12895-018-0081-1.