{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T04:57:27Z","timestamp":1790139447021,"version":"4.0.1"},"reference-count":34,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2022,6,26]],"date-time":"2022-06-26T00:00:00Z","timestamp":1656201600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"S\u00e3o Paulo Research Foundation (FAPESP) to Jos\u00e9 Pontes Jr","award":["#2015\/17097-2"],"award-info":[{"award-number":["#2015\/17097-2"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Penile cancer (PeC) is a rare disease, and no prognostic biomarkers have been adopted in clinical practice yet. The objective of the present study was to identify differentially expressed miRNAs (DEmiRs) and genes (DEGs) as potential biomarkers for lymph node metastasis and other prognostic factors in PeC. Tumor samples were prospectively obtained from 24 patients with squamous cell carcinoma of the penis. miRNA microarray analysis was performed comparing tumors from patients with inguinal lymph node metastatic and localized disease, and the results were validated by qRT-PCR. Eighty-three gene expression levels were also compared between groups through qRT-PCR. Moreover, DEmiRs and DEGs expression levels were correlated with clinicopathological variables, cancer-specific (CSS), and overall survival (OS). TAC software, TM4 MeV 4.9 software, SPSS v.25.0, and R software v.4.0.2 were used for statistical analyses. We identified 21 DEmiRs in microarray analysis, and seven were selected for validation. miR-744-5p and miR-421 were overexpressed in tissue samples of metastatic patients, and high expression of miR-421 was also associated with lower OS. We found seven DEGs (CCND1, EGFR, ENTPD5, HOXA10, IGF1R, MYC, and SNAI2) related to metastatic disease. A significant association was found between increased MMP1 expression and tumor size, grade, pathological T stage, and perineural invasion. Other genes were also associated with clinicopathological variables, CSS and OS. Finally, we found changes in mRNA\u2013miRNA regulation that contribute to understanding the mechanisms involved in tumor progression. Therefore, we identified miRNA and mRNA expression profiles as potential biomarkers associated with lymph node metastasis and prognosis in PeC, in addition to disruption in mRNA\u2013miRNA regulation during disease progression.<\/jats:p>","DOI":"10.3390\/ijms23137103","type":"journal-article","created":{"date-parts":[[2022,6,26]],"date-time":"2022-06-26T09:00:13Z","timestamp":1656234013000},"page":"7103","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["miRNA and mRNA Expression Profiles Associated with Lymph Node Metastasis and Prognosis in Penile Carcinoma"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7813-7128","authenticated-orcid":false,"given":"Claudio B.","family":"Murta","sequence":"first","affiliation":[{"name":"Urology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatiane K.","family":"Furuya","sequence":"additional","affiliation":[{"name":"Center for Translational Research in Oncology (LIM24), Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7372-2608","authenticated-orcid":false,"given":"Alexis G. M.","family":"Carrasco","sequence":"additional","affiliation":[{"name":"Center for Translational Research in Oncology (LIM24), Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miyuki","family":"Uno","sequence":"additional","affiliation":[{"name":"Center for Translational Research in Oncology (LIM24), Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7999-5818","authenticated-orcid":false,"given":"Laura","family":"Sichero","sequence":"additional","affiliation":[{"name":"Center for Translational Research in Oncology (LIM24), Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luisa L.","family":"Villa","sequence":"additional","affiliation":[{"name":"Center for Translational Research in Oncology (LIM24), Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheila F.","family":"Faraj","sequence":"additional","affiliation":[{"name":"Pathology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael F.","family":"Coelho","sequence":"additional","affiliation":[{"name":"Urology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuliano B.","family":"Guglielmetti","sequence":"additional","affiliation":[{"name":"Urology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mauricio D.","family":"Cordeiro","sequence":"additional","affiliation":[{"name":"Urology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katia R. M.","family":"Leite","sequence":"additional","affiliation":[{"name":"Urology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2615-7730","authenticated-orcid":false,"given":"William C.","family":"Nahas","sequence":"additional","affiliation":[{"name":"Urology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0342-8726","authenticated-orcid":false,"given":"Roger","family":"Chammas","sequence":"additional","affiliation":[{"name":"Center for Translational Research in Oncology (LIM24), Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"Jos\u00e9","family":"Pontes","sequence":"additional","affiliation":[{"name":"Urology Department, Instituto do Cancer do Estado de Sao Paulo (ICESP), Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo (HCFMUSP), Sao Paulo CEP 01246-000, SP, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","unstructured":"Hakenberg, O.W., Comp\u00e9rat, E., Minhas, S., Necchi, A., Protzel, C., Watkin, N., and Comp\u00e9rat, E. (2018, June 05). The European Association of Urology (EAU) Guidelines on Penile Cancer. Available online: http:\/\/uroweb.org\/guideline\/penile-cancer\/#10."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"503","DOI":"10.1016\/j.brachy.2019.04.007","article-title":"National patterns of care for early-stage penile cancers in the United States: How is radiation and brachytherapy utilized?","volume":"18","author":"Mulherkar","year":"2019","journal-title":"Brachytherapy"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1111\/bju.13009","article-title":"Predicting postoperative complications of inguinal lymph node dissection for penile cancer in an international multicentre cohort","volume":"116","author":"Gopman","year":"2015","journal-title":"BJU Int."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1002\/jso.20980","article-title":"Surgical treatment of invasive squamous cell carcinoma of the penis: Brazilian National Cancer Institute long-term experience","volume":"97","author":"Ornellas","year":"2008","journal-title":"J. Surg. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1354","DOI":"10.1016\/j.juro.2008.06.028","article-title":"Prognostic factors in invasive squamous cell carcinoma of the penis: Analysis of 196 patients treated at the Brazilian National Cancer Institute","volume":"180","author":"Ornellas","year":"2008","journal-title":"J. Urol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2910","DOI":"10.1002\/cncr.23742","article-title":"Examining the association between socioeconomic status and potential human papillomavirus-associated cancers","volume":"113","author":"Benard","year":"2008","journal-title":"Cancer"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3376","DOI":"10.1016\/j.ajpath.2014.08.004","article-title":"Profiling of expression of human papillomavirus-related cancer miRNAs in penile squamous cell carcinomas","volume":"184","author":"Barzon","year":"2014","journal-title":"Am. J. Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Zhang, L., Wei, P., Shen, X., Zhang, Y., Xu, B., Zhou, J., Fan, S., Hao, Z., Shi, H., and Zhang, X. (2015). MicroRNA Expression Profile in Penile Cancer Revealed by Next-Generation Small RNA Sequencing. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0131336"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1016\/j.juro.2016.01.115","article-title":"Integrated Loss of miR-1\/miR-101\/miR-204 Discriminates Metastatic from Nonmetastatic Penile Carcinomas and Can Predict Patient Outcome","volume":"196","author":"Hartz","year":"2016","journal-title":"J. Urol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"15294","DOI":"10.18632\/oncotarget.14783","article-title":"Integrative miRNA and mRNA analysis in penile carcinomas reveals markers and pathways with potential clinical impact","volume":"8","author":"Kuasne","year":"2017","journal-title":"Oncotarget"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"391","DOI":"10.31557\/APJCP.2020.21.2.391","article-title":"MIR-107, MIR-223-3P and MIR-21-5P Reveals Potential Biomarkers in Penile Cancer","volume":"21","author":"Pinho","year":"2020","journal-title":"Asian Pac. J. Cancer Prev."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Ayoubian, H., Heinzelmann, J., Holters, S., Khalmurzaev, O., Pryalukhin, A., Loertzer, P., Heinzelbecker, J., Lohse, S., Geppert, C., and Loertzer, H. (2021). miRNA Expression Characterizes Histological Subtypes and Metastasis in Penile Squamous Cell Carcinoma. Cancers, 13.","DOI":"10.3390\/cancers13061480"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Furuya, T.K., Murta, C.B., Murillo Carrasco, A.G., Uno, M., Sichero, L., Villa, L.L., Cardilli, L., Coelho, R.F., Guglielmetti, G.B., and Cordeiro, M.D. (2021). Disruption of miRNA-mRNA Networks Defines Novel Molecular Signatures for Penile Carcinogenesis. Cancers, 13.","DOI":"10.3390\/cancers13194745"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2627","DOI":"10.2147\/CMAR.S167432","article-title":"Upregulated expression of miR-421 is associated with poor prognosis in non-small-cell lung cancer","volume":"10","author":"Li","year":"2018","journal-title":"Cancer Manag. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2625","DOI":"10.3892\/etm.2017.4798","article-title":"MicroRNA-421 promotes the proliferation and metastasis of gastric cancer cells by targeting claudin-11","volume":"14","author":"Yang","year":"2017","journal-title":"Exp. Ther. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1465","DOI":"10.1245\/s10434-021-10688-6","article-title":"MiR-744 Functions as an Oncogene Through Direct Binding to c-Fos Promoter and Facilitates Non-small Cell Lung Cancer Progression","volume":"29","author":"Li","year":"2022","journal-title":"Ann. Surg. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1467","DOI":"10.1038\/bjc.2015.366","article-title":"Plasma microRNA profiles: Identification of miR-744 as a novel diagnostic and prognostic biomarker in pancreatic cancer","volume":"113","author":"Miyamae","year":"2015","journal-title":"Br. J. Cancer"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5219","DOI":"10.1158\/0008-5472.CAN-15-1004","article-title":"Genomic Profiling of Penile Squamous Cell Carcinoma Reveals New Opportunities for Targeted Therapy","volume":"75","author":"McDaniel","year":"2015","journal-title":"Cancer Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1634\/theoncologist.2015-0241","article-title":"Comprehensive Genomic Profiling of Advanced Penile Carcinoma Suggests a High Frequency of Clinically Relevant Genomic Alterations","volume":"21","author":"Ali","year":"2016","journal-title":"Oncologist"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1111\/bju.12450","article-title":"Epidermal growth factor receptor-targeted therapy in locally advanced or metastatic squamous cell carcinoma of the penis","volume":"113","author":"Carthon","year":"2014","journal-title":"BJU Int."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Loh, C.-Y., Chai, J., Tang, T., Wong, W., Sethi, G., Shanmugam, M., Chong, P., and Looi, C. (2019). The E-Cadherin and N-Cadherin Switch in Epithelial-to-Mesenchymal Transition: Signaling, Therapeutic Implications, and Challenges. Cells, 8.","DOI":"10.3390\/cells8101118"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2249","DOI":"10.1002\/ijc.26226","article-title":"Deregulation of Snai2 is associated with metastasis and poor prognosis in tongue squamous cell carcinoma","volume":"130","author":"Wang","year":"2012","journal-title":"Int. J. Cancer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"8437","DOI":"10.2147\/OTT.S221340","article-title":"The Role Of PD-1\/PD-L1 Axis In Treg Development And Function: Implications For Cancer Immunotherapy","volume":"12","author":"Cai","year":"2019","journal-title":"Onco Targets Ther."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.euo.2018.07.010","article-title":"PD-L1 Expression in Men with Penile Cancer and its Association with Clinical Outcomes","volume":"2","author":"Davidsson","year":"2019","journal-title":"Eur. Urol. Oncol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"97","DOI":"10.12703\/P6-97","article-title":"MyD88: A central player in innate immune signaling","volume":"6","author":"Deguine","year":"2014","journal-title":"F1000Prime Rep."},{"key":"ref_27","first-page":"5515","article-title":"Down-regulation of Toll-like Receptor TLR4 Is Associated with HPV DNA Integration in Penile Carcinoma","volume":"37","author":"Damasdi","year":"2017","journal-title":"Anticancer Res."},{"key":"ref_28","first-page":"15128","article-title":"Correlation between microRNA-421 expression level and prognosis of gastric cancer","volume":"8","author":"Liu","year":"2015","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"176","DOI":"10.1038\/13802","article-title":"Genomic instability in Gadd45a-deficient mice","volume":"23","author":"Hollander","year":"1999","journal-title":"Nat. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9671","DOI":"10.1158\/0008-5472.CAN-10-2177","article-title":"Gadd45a Functions as a Promoter or Suppressor of Breast Cancer Dependent on the Oncogenic Stress","volume":"70","author":"Tront","year":"2010","journal-title":"Cancer Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5245","DOI":"10.1158\/0008-5472.CAN-04-0496","article-title":"Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets","volume":"64","author":"Andersen","year":"2004","journal-title":"Cancer Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1126\/science.2448875","article-title":"Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase","volume":"239","author":"Saiki","year":"1988","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2508","DOI":"10.1128\/JCM.37.8.2508-2517.1999","article-title":"Development and clinical evaluation of a highly sensitive PCR-reverse hybridization line probe assay for detection and identification of anogenital human papillomavirus","volume":"37","author":"Kleter","year":"1999","journal-title":"J. Clin. Microbiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/13\/7103\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:38:41Z","timestamp":1760139521000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/13\/7103"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,26]]},"references-count":34,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["ijms23137103"],"URL":"https:\/\/doi.org\/10.3390\/ijms23137103","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,26]]}}}