{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:34:08Z","timestamp":1760236448401,"version":"build-2065373602"},"reference-count":42,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,11,30]],"date-time":"2021-11-30T00:00:00Z","timestamp":1638230400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100012000","name":"Fundaci\u00f3n CajaCanarias","doi-asserted-by":"publisher","award":["Salucan 13"],"award-info":[{"award-number":["Salucan 13"]}],"id":[{"id":"10.13039\/100012000","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014851","name":"Fundaci\u00f3n DISA","doi-asserted-by":"publisher","award":["DISA01"],"award-info":[{"award-number":["DISA01"]}],"id":[{"id":"10.13039\/501100014851","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CIMB"],"abstract":"<jats:p>Formalin-fixed paraffin-embedded (FFPE) tumour samples may provide crucial data regarding biomarkers for neoplasm progression. Analysis of gene expression is frequently used for this purpose. Therefore, mRNA expression needs to be normalized through comparison to reference genes. In this study, we establish which of the usually reported reference genes is the most reliable one in cutaneous malignant melanoma (MM) and cutaneous squamous cell carcinoma (CSCC). ACTB, TFRC, HPRT1 and TBP expression was quantified in 123 FFPE samples (74 MM and 49 CSCC biopsies) using qPCR. Expression stability was analysed by NormFinder and Bestkeeper softwares, and the direct comparison method between means and SD. The in-silico analysis with BestKeeper indicated that HPRT1 was more stable than ACTB and TFRC in MM (1.85 vs. 2.15) and CSCC tissues (2.09 vs. 2.33). The best option to NormFinder was ACTB gene (0.56) in MM and TFRC (0.26) in CSCC. The direct comparison method showed lower SD means of ACTB expression in MM (1.17) and TFRC expression in CSCC samples (1.00). When analysing the combination of two reference genes for improving stability, NormFinder indicated HPRT1 and ACTB to be the best for MM samples, and HPRT1 and TFRC genes for CSCC. In conclusion, HPRT1 and ACTB genes in combination are the most appropriate choice for normalization in gene expression studies in MM FFPE tissue, while the combination of HPRT1 and TFRC genes are the best option in analysing CSCC FFPE samples. These may be used consistently in forthcoming studies on gene expression in both tumours.<\/jats:p>","DOI":"10.3390\/cimb43030151","type":"journal-article","created":{"date-parts":[[2021,12,1]],"date-time":"2021-12-01T05:03:16Z","timestamp":1638334996000},"page":"2167-2176","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Gene Expression Studies in Formalin-Fixed Paraffin-Embedded Samples of Cutaneous Cancer: The Need for Reference Genes"],"prefix":"10.3390","volume":"43","author":[{"given":"Omar","family":"Garc\u00eda-P\u00e9rez","sequence":"first","affiliation":[{"name":"Research Unit, Hospital Universitario Nuestra Se\u00f1ora de Candelaria, Ctra. Gral. del Rosario, 145, 38010 Santa Cruz de Tenerife, Spain"},{"name":"Universidad de La Laguna, Calle Padre Herrera, s\/n, 38200 San Crist\u00f3bal de La Laguna, Spain"},{"name":"Instituto Universitario de Enfermedades Tropicales y Salud P\u00fablica de Canarias (IUETSPC), 38296 San Crist\u00f3bal de La Laguna, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leticia","family":"Melgar-Vilaplana","sequence":"additional","affiliation":[{"name":"Pathology Department, Hospital Universitario Nuestra Se\u00f1ora de Candelaria, Ctra. Gral. del Rosario, 145, 38010 Santa Cruz de Tenerife, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5534-8280","authenticated-orcid":false,"given":"Elizabeth","family":"C\u00f3rdoba-Lan\u00fas","sequence":"additional","affiliation":[{"name":"Research Unit, Hospital Universitario Nuestra Se\u00f1ora de Candelaria, Ctra. Gral. del Rosario, 145, 38010 Santa Cruz de Tenerife, Spain"},{"name":"Universidad de La Laguna, Calle Padre Herrera, s\/n, 38200 San Crist\u00f3bal de La Laguna, Spain"},{"name":"Instituto Universitario de Enfermedades Tropicales y Salud P\u00fablica de Canarias (IUETSPC), 38296 San Crist\u00f3bal de La Laguna, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4120-9962","authenticated-orcid":false,"given":"Ricardo","family":"Fern\u00e1ndez-de-Misa","sequence":"additional","affiliation":[{"name":"Research Unit, Hospital Universitario Nuestra Se\u00f1ora de Candelaria, Ctra. Gral. del Rosario, 145, 38010 Santa Cruz de Tenerife, Spain"},{"name":"Universidad de La Laguna, Calle Padre Herrera, s\/n, 38200 San Crist\u00f3bal de La Laguna, Spain"},{"name":"Dermatology Department, Hospital Universitario Nuestra Se\u00f1ora de Candelaria, Ctra. Gral. del Rosario, 145, 38010 Santa Cruz de Tenerife, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,11,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.suc.2019.09.005","article-title":"Epidemiology and Risk Factors of Melanoma","volume":"100","author":"Carr","year":"2019","journal-title":"Surg. Clin. N. Am."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"400","DOI":"10.7150\/jca.21381","article-title":"Identification of Biomarker for Cutaneous Squamous Cell Carcinoma Using Microarray Data Analysis","volume":"9","author":"Wei","year":"2018","journal-title":"J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Belter, B., Haase-Kohn, C., and Pietzsch, J. (2017). Biomarkers in Malignant Melanoma: Recent Trends and Critical Perspective. 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