{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T06:50:02Z","timestamp":1771051802558,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2019,12,8]],"date-time":"2019-12-08T00:00:00Z","timestamp":1575763200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The current development of BRAF inhibitors has revolutionized the treatment of unresectable melanoma. As the potential heterogeneity of BRAF mutations in melanoma has been reported, accurate detection of BRAF mutations are important. However, the genetic heterogeneity of acral melanoma\u2014a distinct type of melanoma with a unique genetic background\u2014has not fully been investigated. We conducted a retrospective review of our acral melanoma patients. Of the 196 patients with acral melanoma, we retrieved 31 pairs of primary and matched metastatic melanomas. We immunostained the 31 pairs with VE1, a BRAFV600E-mutation-specific monoclonal antibody. Immunohistochemistry with VE1 showed a high degree of sensitivity and specificity for detecting BRAFV600E mutations compared with the real-time polymerase chain reaction method. A total of nine primary (29.0%) and eight metastatic (25.8%) acral melanomas were positive for VE1. In three patients (9.7%), we observed a discordance of VE1 staining between the primary and metastatic lesions. Of note, VE1 immunohistochemical staining revealed a remarkable degree of intra-tumor genetic heterogeneity in acral melanoma. Our study reveals that VE1 immunostaining is a useful ancillary method for detecting BRAFV600E mutations in acral melanoma and allows for a clear visualization of intra- and inter-tumor BRAF heterogeneity.<\/jats:p>","DOI":"10.3390\/ijms20246191","type":"journal-article","created":{"date-parts":[[2019,12,9]],"date-time":"2019-12-09T05:54:51Z","timestamp":1575870891000},"page":"6191","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Intra- and Inter-Tumor BRAF Heterogeneity in Acral Melanoma: An Immunohistochemical Analysis"],"prefix":"10.3390","volume":"20","author":[{"given":"Takamichi","family":"Ito","sequence":"first","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yumiko","family":"Kaku-Ito","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1068-6873","authenticated-orcid":false,"given":"Maho","family":"Murata","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshio","family":"Ichiki","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"},{"name":"Department of Anatomic Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuki","family":"Kuma","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"},{"name":"Department of Anatomic Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8433-6035","authenticated-orcid":false,"given":"Yuka","family":"Tanaka","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Taketoshi","family":"Ide","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fumitaka","family":"Ohno","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maiko","family":"Wada-Ohno","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuichi","family":"Yamada","sequence":"additional","affiliation":[{"name":"Department of Anatomic Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshinao","family":"Oda","sequence":"additional","affiliation":[{"name":"Department of Anatomic Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2967-1073","authenticated-orcid":false,"given":"Masutaka","family":"Furue","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,12,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"212","DOI":"10.3322\/caac.20121","article-title":"Cancer statistics, 2011: The impact of eliminating socioeconomic and racial disparities on premature cancer deaths","volume":"61","author":"Siegel","year":"2011","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1093\/jnci\/djr077","article-title":"Annual report to the nation on the status of cancer, 1975\u20132007, featuring tumors of the brain and other nervous system","volume":"103","author":"Kohler","year":"2011","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1056\/NEJMoa1112302","article-title":"Survival in BRAF V600-mutant advanced melanoma treated with vemurafenib","volume":"366","author":"Sosman","year":"2012","journal-title":"N. Engl. J. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1056\/NEJMoa1203421","article-title":"Improved survival with MEK inhibition in BRAF-mutated melanoma","volume":"367","author":"Flaherty","year":"2012","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1016\/S0140-6736(12)60868-X","article-title":"Dabrafenib in BRAF-mutated metastatic melanoma: A multicentre, open-label, phase 3 randomised controlled trial","volume":"380","author":"Hauschild","year":"2012","journal-title":"Lancet"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2507","DOI":"10.1056\/NEJMoa1103782","article-title":"Improved survival with vemurafenib in melanoma with BRAF V600E mutation","volume":"364","author":"Chapman","year":"2011","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1038\/nature00766","article-title":"Mutations of the BRAF gene in human cancer","volume":"417","author":"Davies","year":"2002","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1097\/CCO.0b013e3282f5271c","article-title":"The MAPK pathway in melanoma","volume":"20","author":"Fecher","year":"2008","journal-title":"Curr. Opin. Oncol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"367","DOI":"10.6004\/jnccn.2019.0018","article-title":"Cutaneous melanoma, version 2.2019, NCCN Clinical Practice Guidelines in Oncology","volume":"17","author":"Coit","year":"2019","journal-title":"J. Natl. Compr. Cancer Netw."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.sder.2012.07.007","article-title":"Molecular platforms utilized to detect BRAF V600E mutation in melanoma","volume":"31","author":"Curry","year":"2012","journal-title":"Semin. Cutan. Med. Surg."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Ihle, M.A., Fassunke, J., K\u00f6nig, K., Gr\u00fcnewald, I., Schlaak, M., Kreuzberg, N., Tietze, L., Schildhaus, H.U., B\u00fcttner, R., and Merkelbach-Bruse, S. (2014). Comparison of high resolution melting analysis, pyrosequencing, next generation sequencing and immunohistochemistry to conventional Sanger sequencing for the detection of p.V600E and non-p.V600E BRAF mutations. BMC Cancer, 14.","DOI":"10.1186\/1471-2407-14-13"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1016\/j.jaad.2012.03.022","article-title":"Detection of BRAF V600E mutations in skin metastases of malignant melanoma by monoclonal antibody VE1","volume":"67","author":"Skorokhod","year":"2012","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1007\/s00401-011-0841-z","article-title":"Assessment of BRAF V600E mutation status by immunohistochemistry with a mutation-specific monoclonal antibody","volume":"122","author":"Capper","year":"2011","journal-title":"Acta Neuropathol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1038\/nature12624","article-title":"Tumour heterogeneity and cancer cell plasticity","volume":"501","author":"Meacham","year":"2013","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Yancovitz, M., Litterman, A., Yoon, J., Ng, E., Shapiro, R.L., Berman, R.S., Pavlick, A.C., Darvishian, F., Christos, P., and Mazumdar, M. (2012). Intra- and inter-tumor heterogeneity of BRAF(V600E) mutations [sic] in primary and metastatic melanoma. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0029336"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1038\/sj.bjc.6606072","article-title":"Polyclonality of BRAF mutations in primary melanoma and the selection of mutant alleles during progression","volume":"104","author":"Lin","year":"2011","journal-title":"Br. J. Cancer"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.ejca.2017.05.015","article-title":"Discrepancy in BRAF status among patients with metastatic malignant melanoma: A meta-analysis","volume":"81","author":"Valachis","year":"2017","journal-title":"Eur. J. Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e285","DOI":"10.1097\/MD.0000000000000285","article-title":"BRAFV600E immunopositive melanomas show low frequency of heterogeneity and association with epithelioid tumor cells: A STROBE-compliant article","volume":"93","author":"Verlinden","year":"2014","journal-title":"Medicine (Baltimore)"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1111\/1346-8138.13834","article-title":"Acral lentiginous melanoma versus other melanoma: A single-center analysis in Japan","volume":"44","author":"Wada","year":"2017","journal-title":"J. Dermatol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.jaad.2014.10.008","article-title":"Acral lentiginous melanoma: Who benefits from sentinel lymph node biopsy?","volume":"72","author":"Ito","year":"2015","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"912","DOI":"10.1111\/jdv.12722","article-title":"Triple-marker PCR assay of sentinel lymph node as a prognostic factor in melanoma","volume":"29","author":"Ito","year":"2015","journal-title":"J. Eur. Acad. Dermatol. Venereol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.jdermsci.2015.07.012","article-title":"Clinical characteristics associated with BRAF, NRAS and KIT mutations in Japanese melanoma patients","volume":"80","author":"Sakaizawa","year":"2015","journal-title":"J. Dermatol. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2404","DOI":"10.1056\/NEJMc1512354","article-title":"Melanomas and mechanical stress points on the plantar surface of the foot","volume":"374","author":"Minagawa","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1007\/s11864-019-0666-x","article-title":"Adjuvant herapy for nelanoma","volume":"20","author":"Ito","year":"2019","journal-title":"Curr. Treat. Options Oncol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1007\/s11912-018-0676-z","article-title":"Melanoma and immune checkpoint inhibitors","volume":"20","author":"Furue","year":"2018","journal-title":"Curr. Oncol. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1097\/DAD.0000000000000404","article-title":"BRAF-V600 mutation heterogeneity in primary and metastatic melanoma: A study with pyrosequencing and immunohistochemistry","volume":"2","author":"Yaman","year":"2016","journal-title":"Am. J. Dermatopathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"630","DOI":"10.2340\/00015555-2326","article-title":"Highly concordant results between immunohistochemistry and molecular testing of mutated V600E BRAF in primary and metastatic melanoma","volume":"96","author":"Manfredi","year":"2016","journal-title":"Acta Derm. Venereol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1001\/jamadermatol.2014.3689","article-title":"BRAFV600E protein expression in primary cutaneous malignant melanomas and paired metastases","volume":"151","author":"Eriksson","year":"2015","journal-title":"JAMA Dermatol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3078","DOI":"10.1038\/jid.2015.229","article-title":"Intra- and inter-tumoral homogeneity of BRAF(V600E) mutations in melanoma tumors","volume":"135","author":"Villanueva","year":"2015","journal-title":"J. Investig. Dermatol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.humpath.2014.08.018","article-title":"Prospective immunohistochemical analysis of BRAF V600E mutation in melanoma","volume":"46","author":"Thiel","year":"2015","journal-title":"Hum. Pathol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1016\/j.humpath.2014.07.014","article-title":"Accurate detection of BRAF p.V600E mutations in challenging melanoma specimens requires stringent immunohistochemistry scoring criteria or sensitive molecular assays","volume":"45","author":"Fisher","year":"2014","journal-title":"Hum. Pathol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/j.yexmp.2014.09.008","article-title":"Comparing BRAF mutation status in matched primary and metastatic cutaneous melanomas: Implications on optimized targeted therapy","volume":"97","author":"Saroufim","year":"2014","journal-title":"Exp. Mol. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1097\/PAS.0000000000000136","article-title":"Intrapatient homogeneity of BRAFV600E expression in melanoma","volume":"38","author":"Menzies","year":"2014","journal-title":"Am. J. Surg. Pathol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Boursault, L., Haddad, V., Vergier, B., Cappellen, D., Verdon, S., Bellocq, J.P., Jouary, T., and Merlio, J.P. (2013). Tumor homogeneity between primary and metastatic sites for BRAF status in metastatic melanoma determined by immunohistochemical and molecular testing. PLoS ONE., 8.","DOI":"10.1371\/journal.pone.0070826"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.jdermsci.2016.10.006","article-title":"Comparative study on driver mutations in primary and metastatic melanomas at a single Japanese institute: A clue for intra- and inter-tumor heterogeneity","volume":"85","author":"Kaji","year":"2017","journal-title":"J. Dermatol. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1016\/j.jdermsci.2013.07.013","article-title":"KIT, NRAS, BRAF and PTEN mutations in a sample of Swedish patients with acral lentiginous melanoma","volume":"72","author":"Zebary","year":"2013","journal-title":"J. Dermatol. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1097\/PAS.0b013e318271249e","article-title":"Immunohistochemical analysis of BRAF(V600E) expression of primary and metastatic melanoma and comparison with mutation status and melanocyte differentiation antigens of metastatic lesions","volume":"37","author":"Busam","year":"2013","journal-title":"Am. J. Surg. Pathol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1097\/PAS.0b013e31826485c0","article-title":"Immunohistochemistry is highly sensitive and specific for the detection of V600E BRAF mutation in melanoma","volume":"37","author":"Long","year":"2013","journal-title":"Am. J. Surg. Pathol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.jmoldx.2012.09.001","article-title":"Detection of BRAF p.V600E mutations in melanomas: Comparison of four methods argues for sequential use of immunohistochemistry and pyrosequencing","volume":"15","author":"Colomba","year":"2013","journal-title":"J. Mol. Diagn."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"823","DOI":"10.7150\/jca.14461","article-title":"Prognostic significance of forkhead box M1 (FOXM1) expression and antitumor effect of FOXM1 inhibition in angiosarcoma","volume":"7","author":"Ito","year":"2016","journal-title":"J. Cancer"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1111\/his.12909","article-title":"Prognostic significance of forkhead box M1 (FoxM1) expression and antitumour effect of FoxM1 inhibition in melanoma","volume":"69","author":"Ito","year":"2016","journal-title":"Histopathology"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1036","DOI":"10.1016\/j.ajpath.2015.12.013","article-title":"Activation of the OVOL1-OVOL2 axis in the hair bulb and in pilomatricoma","volume":"186","author":"Ito","year":"2016","journal-title":"Am. J. Pathol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1038\/modpathol.2016.169","article-title":"Potential role of the OVOL1-OVOL2 axis and c-Myc in the progression of cutaneous squamous cell carcinoma","volume":"30","author":"Ito","year":"2017","journal-title":"Mod. Pathol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/24\/6191\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:40:25Z","timestamp":1760190025000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/24\/6191"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,12,8]]},"references-count":43,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["ijms20246191"],"URL":"https:\/\/doi.org\/10.3390\/ijms20246191","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,12,8]]}}}