BRAF V600E Mutations and Beyond: A Molecular Perspective of Melanoma from a Tertiary Cancer Referral Center of India

 

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Abstract

Objectives Malignant melanoma demonstrates frequently occurring mutations of genes in the serine/threonine kinase pathway, namely BRAF, NRAS, and neurofibromin 1. There is rare documentation of a detailed analysis of these mutations in cases of melanoma among Indian patients. We present molecular features in cases of malignant melanoma, diagnosed at a tertiary cancer referral center in India, over a period of 8 years (2011–2018).

Materials and Methods This study was performed on formalin fixed paraffin embedded tissues of 88 histologically confirmed cases of malignant melanoma. BRAF gene alterations were studied by both Sanger sequencing and real-time polymerase chain reaction techniques (n = 74). Molecular testing for BRAF and NRAS gene alterations was accomplished in 74/88 cases (80%). Molecular test results were correlated with clinicopathological features using IBM SPSS Statistical software 25.0.

Results The age ranged from 13 to 79 years (median = 57), with a M:F ratio of 1.4:1. BRAF mutations were observed in 12/74 (16.21%) patients, including V600E (n = 7), A594T (n = 1), T599 = (n = 2), V600K (n = 1), and Q612P (n = 1), while NRAS mutations were observed in 6/38 (15.7%) patients. Among various subtypes, nodular melanoma was the most frequent subtype (33%) among cutaneous malignant melanomas. Among non-cutaneous melanomas, mucosal melanomas were observed in 37.5% of cases.

Conclusion This constitutes one of the few reports on comprehensive analysis of molecular alterations underlying melanomas in Indian patients. A larger sample size, with more extensive molecular markers, would yield additional information on the disease manifestation.

Authors' Contributions

O.S. contributed to conceptualization, methodology, administration. V.V. helped in data curation, writing-original draft preparation. M.G. contributed to methodology and investigation. P.B. helped in investigation and supervision. N.K. contributed to analysis and investigation. G.W. and M.R. were involved in investigation. B.R. and E.S. reviewed and edited the manuscript. S.D. helped in administration and visualization.

Publication History

Article published online:
02 March 2023

© 2023. MedIntel Services Pvt Ltd. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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• References

• 1 Kang X, Zeng Y, Liang J. et al. Aberrations and clinical significance of BRAF in malignant melanoma: a series of 60 cases in Chinese Uyghur. Medicine (Baltimore) 2018; 97 (01) e9509
  CrossrefPubMedGoogle Scholar
• 2 Grzywa TM, Paskal W, Włodarski PK. Intratumor and intertumor heterogeneity in melanoma. Transl Oncol 2017; 10 (06) 956-975
  CrossrefPubMedGoogle Scholar
• 3 Cheng L, Lopez-Beltran A, Massari F, MacLennan GT, Montironi R. Molecular testing for BRAF mutations to inform melanoma treatment decisions: a move toward precision medicine. Mod Pathol 2018; 31 (01) 24-38
  CrossrefPubMedGoogle Scholar
• 4 Nissan MH, Pratilas CA, Jones AM. et al. Loss of NF1 in cutaneous melanoma is associated with RAS activation and MEK dependence. Cancer Res 2014; 74 (08) 2340-2350
  CrossrefPubMedGoogle Scholar
• 5 Shaughnessy M, Klebanov N, Tsao H. Clinical and therapeutic implications of melanoma genomics. J Transl Genet Genom 2018; 2: 14-26
  Google Scholar
• 6 Bhatia P, Friedlander P, Zakaria EA, Kandil E. Impact of BRAF mutation status in the prognosis of cutaneous melanoma: an area of ongoing research. [published correction appears in Ann Transl Med. 2015 Mar;3(4):60] Ann Transl Med 2015; 3 (02) 24
  PubMedGoogle Scholar
• 7 Heppt MV, Siepmann T, Engel J. et al. Prognostic significance of BRAF and NRAS mutations in melanoma: a German study from routine care. BMC Cancer 2017; 17 (01) 536
  CrossrefPubMedGoogle Scholar
• 8 Rutkowski P, Gos A, Jurkowska M. et al. Molecular alterations in clinical stage III cutaneous melanoma: correlation with clinicopathological features and patient outcome. Oncol Lett 2014; 8 (01) 47-54
  CrossrefPubMedGoogle Scholar
• 9 Edlundh-Rose E, Egyházi S, Omholt K. et al. NRAS and BRAF mutations in melanoma tumours in relation to clinical characteristics: a study based on mutation screening by pyrosequencing. Melanoma Res 2006; 16 (06) 471-478
  CrossrefPubMedGoogle Scholar
• 10 Si L, Kong Y, Xu X. et al. Prevalence of BRAF V600E mutation in Chinese melanoma patients: large scale analysis of BRAF and NRAS mutations in a 432-case cohort. Eur J Cancer 2012; 48 (01) 94-100
  CrossrefPubMedGoogle Scholar
• 11 Spathis A, Katoulis AC, Damaskou V. et al. BRAF mutation status in primary, recurrent, and metastatic malignant melanoma and its relation to histopathological parameters. Dermatol Pract Concept 2019; 9 (01) 54-62
  PubMedGoogle Scholar
• 12 Ugurel S, Thirumaran RK, Bloethner S. et al. B-RAF and N-RAS mutations are preserved during short time in vitro propagation and differentially impact prognosis. PLoS One 2007; 2 (02) e236
  CrossrefPubMedGoogle Scholar
• 13 Ferlay J, Ervik M, Lam F, Colombet M, Mery L, Piñeros M, Znaor A, Soerjomataram I, Bray F. (2020). Global Cancer Observatory: Cancer Today. Lyon, France: International Agency for Research on Cancer. Accessed January 04, 2023, from: https://gco.iarc.fr/today
  Google Scholar
• 14 Badwe RA, Pramesh CS, Ganesh B. Hospital based Cancer Registry of Tata Memorial Hospital-2017. Published in 2019
  Google Scholar
• 15 Chang JW, Guo J, Hung CY. et al. Sunrise in melanoma management: time to focus on melanoma burden in Asia. Asia Pac J Clin Oncol 2017; 13 (06) 423-427
  CrossrefPubMedGoogle Scholar
• 16 Panda S, Dash S, Besra K, Samantaray S, Pathy PC, Rout N. Clinicopathological study of malignant melanoma in a regional cancer center. Indian J Cancer 2018; 55 (03) 292-296
  CrossrefPubMedGoogle Scholar
• 17 Radhika K, Prayaga AK, Sundaram C. A clinicopathologic study of malignant melanoma based on cytomorphology. Indian J Cancer 2016; 53 (01) 199-203
  CrossrefPubMedGoogle Scholar
• 18 Sharma K, Mohanti BK, Rath GK. Malignant melanoma: a retrospective series from a regional cancer center in India. J Cancer Res Ther 2009; 5 (03) 173-180
  CrossrefPubMedGoogle Scholar
• 19 Ahmad F, Avabhrath N, Natarajan S, Parikh J, Patole K, Das BR. Molecular evaluation of BRAF V600 mutation and its association with clinicopathological characteristics: first findings from Indian malignant melanoma patients. Cancer Genet 2019; 231-232: 46-53
  CrossrefPubMedGoogle Scholar
• 20 Lal ST, Banipal RP, Bhatti DJ, Yadav HP. Changing trends of skin cancer: a tertiary care hospital study in Malwa Region of Punjab. J Clin Diagn Res 2016; 10 (06) PC12-PC15
  PubMedGoogle Scholar
• 21 Lasota J, Kowalik A, Wasag B. et al. Detection of the BRAF V600E mutation in colon carcinoma: critical evaluation of the imunohistochemical approach. Am J Surg Pathol 2014; 38 (09) 1235-1241
  CrossrefPubMedGoogle Scholar
• 22 Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 2010; 127 (12) 2893-2917
  CrossrefPubMedGoogle Scholar
• 23 Broekaert SM, Roy R, Okamoto I. et al. Genetic and morphologic features for melanoma classification. Pigment Cell Melanoma Res 2010; 23 (06) 763-770
  CrossrefPubMedGoogle Scholar
• 24 Libra M, Malaponte G, Navolanic PM. et al. Analysis of BRAF mutation in primary and metastatic melanoma. Cell Cycle 2005; 4 (10) 1382-1384
  CrossrefPubMedGoogle Scholar
• 25 Stang A, Parkin DM, Ferlay J, Jöckel KH. International uveal melanoma incidence trends in view of a decreasing proportion of morphological verification. Int J Cancer 2005; 114 (01) 114-123
  CrossrefPubMedGoogle Scholar
• 26 Ohtsuka H, Nagamatsu S. Changing trends in numbers of deaths from malignant melanoma in Japan, 1955-2000. Dermatology 2003; 207 (02) 162-165
  CrossrefPubMedGoogle Scholar
• 27 Lakhtakia R, Mehta A, Nema SK. Melanoma: a frequently missed diagnosis. Med J Armed Forces India 2009; 65 (03) 292-294
  CrossrefPubMedGoogle Scholar
• 28 Sheen YS, Liao YH, Liau JY. et al. Prevalence of BRAF and NRAS mutations in cutaneous melanoma patients in Taiwan. J Formos Med Assoc 2016; 115 (02) 121-127
  CrossrefPubMedGoogle Scholar
• 29 Shim JH, Shin HT, Park J. et al. Mutational profiling of acral melanomas in Korean populations. Exp Dermatol 2017; 26 (10) 883-888
  CrossrefPubMedGoogle Scholar
• 30 Rinonce HT, Aji RPM, Hayati N, Pudjohartono MF, Kameswari B. Irianiwati. Low BRAF V600 mutation prevalence in primary skin nodular melanoma in Indonesia: a real-time PCR detection among Javanese patients. BMC Proc 2019; 13 (Suppl. 11) 15
  CrossrefPubMedGoogle Scholar
• 31 Yamazaki N, Kiyohara Y, Uhara H. et al. Efficacy and safety of nivolumab in Japanese patients with previously untreated advanced melanoma: a phase II study. Cancer Sci 2017; 108 (06) 1223-1230
  CrossrefPubMedGoogle Scholar
• 32 Aksenenko MB, Kirichenko AK, Ruksha TG. Russian study of morphological prognostic factors characterization in BRAF-mutant cutaneous melanoma. Pathol Res Pract 2015; 211 (07) 521-527
  CrossrefPubMedGoogle Scholar
• 33 Gutiérrez-Castañeda LD, Nova JA, Tovar-Parra JD. Frequency of mutations in BRAF, NRAS, and KIT in different populations and histological subtypes of melanoma: a systemic review. Melanoma Res 2020; 30 (01) 62-70
  CrossrefPubMedGoogle Scholar
• 34 Devitt B, Liu W, Salemi R. et al. Clinical outcome and pathological features associated with NRAS mutation in cutaneous melanoma. Pigment Cell Melanoma Res 2011; 24 (04) 666-672
  CrossrefPubMedGoogle Scholar
• 35 Pracht M, Mogha A, Lespagnol A. et al. Prognostic and predictive values of oncogenic BRAF, NRAS, c-KIT and MITF in cutaneous and mucous melanoma. J Eur Acad Dermatol Venereol 2015; 29 (08) 1530-1538
  CrossrefPubMedGoogle Scholar
• 36 Lee JH, Choi JW, Kim YS. Frequencies of BRAF and NRAS mutations are different in histological types and sites of origin of cutaneous melanoma: a meta-analysis. Br J Dermatol 2011; 164 (04) 776-784
  CrossrefPubMedGoogle Scholar
• 37 WHO | Global disease burden from solar ultraviolet radiation - archived, 11 December 2009., WHO [Internet].. 2017 [cited 2020 Sep 6]. Accessed January 4, 2022, at: http://www.who.int/uv/resources/archives/fs305/en/
  PubMed
• 38 Al-Jamal M, Griffith JL, Lim HW. Photoprotection in ethnic skin. Dermatologica Sinica 2014; 32: 217-224
  CrossrefGoogle Scholar
• 39 Saldanha G, Potter L, Daforno P, Pringle JH. Cutaneous melanoma subtypes show different BRAF and NRAS mutation frequencies. Clin Cancer Res 2006; 12 (15) 4499-4505
  CrossrefPubMedGoogle Scholar
• 40 Oyama S, Funasaka Y, Watanabe A, Takizawa T, Kawana S, Saeki H. BRAF, KIT and NRAS mutations and expression of c-KIT, phosphorylated extracellular signal-regulated kinase and phosphorylated AKT in Japanese melanoma patients. J Dermatol 2015; 42 (05) 477-484
  CrossrefPubMedGoogle Scholar
• 41 Carlino MS, Haydu LE, Kakavand H. et al. Correlation of BRAF and NRAS mutation status with outcome, site of distant metastasis and response to chemotherapy in metastatic melanoma. Br J Cancer 2014; 111 (02) 292-299
  CrossrefPubMedGoogle Scholar
• 42 Kulkarni A, Al-Hraishawi H, Simhadri S. et al. BRAF fusion as a novel mechanism of acquired resistance to vemurafenib in BRAF ^V600E mutant melanoma. Clin Cancer Res 2017; 23 (18) 5631-5638
  CrossrefPubMedGoogle Scholar
• 43 Botton T, Talevich E, Mishra VK. et al. Genetic heterogeneity of BRAF fusion kinases in melanoma affects drug responses. Cell Rep 2019; 29 (03) 573-588.e7
  CrossrefPubMedGoogle Scholar
• 44 Busam KJ, Hedvat C, Pulitzer M, von Deimling A, Jungbluth AA. Immunohistochemical analysis of BRAF(V600E) expression of primary and metastatic melanoma and comparison with mutation status and melanocyte differentiation antigens of metastatic lesions. Am J Surg Pathol 2013; 37 (03) 413-420
  CrossrefPubMedGoogle Scholar
• 45 Huang WK, Kuo TT, Wu CE. et al. A comparison of immunohistochemical and molecular methods used for analyzing the BRAF V600E gene mutation in malignant melanoma in Taiwan. Asia Pac J Clin Oncol 2016; 12 (04) 403-408
  CrossrefPubMedGoogle Scholar

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