Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua

Detalhes bibliográficos
Autor(a) principal: Có, Anna Clara Gregório
Data de Publicação: 2023
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
Texto Completo: http://repositorio.ufes.br/handle/10/12554
Resumo: Oral squamous cell carcinoma (OSCC) is the 16th most commonly diagnosed form of cancer globally, with a higher prevalence in the tongue compared to other areas of the oral cavity. However, the lack of effective biomarkers for diagnosis, especially for precancerous lesions, poses a limitation, as visual or histological examination cannot predict the progression of dysplastic lesions, making it difficult to determine whether they will develop into cancer or return to normal epithelium. In this context, the present research aims to investigate molecular targets that may indicate the irreversible transformation of these cells, to provide a basis for broader studies aimed at using these targets as biomarkers for early OSCC diagnosis. To achieve this goal, this experimental study addressed the evaluation of the expression of a panel of microRNAs and proteins in tumour tissues and adjacent tumour-adjacent epithelium obtained from patients with tongue squamous cell carcinoma and oral epithelium from healthy individuals. Additionally, the research explored the association between these biomarkers, seeking to determine their potential application as diagnostic biomarkers for tongue squamous cell carcinoma. A total of 75 cases of tongue squamous cell carcinoma and adjacent tumouradjacent epithelium were included in the study, and the expression of the proteins survivin, Bcl-2, PLK1, p16, p40, p63, EGFR, and cyclin D1 was analysed by immunohistochemistry. The analysis of the microRNA panel's expression involved 31 samples of tongue squamous cell carcinoma, 10 samples of healthy gingival tissue, and 10 samples of serum from healthy individuals, as well as 7 samples of serum from patients diagnosed with OSCC, using the RT-qPCR technique. In silico analysis by bioinformatics validated the findings related to the expression of differentially expressed microRNAs in the sample group. The results showed differences in the expression of miRNA-31-5p (p<0.001) and miRNA-21-5p (p=0.001) in tumour samples compared to control samples. Significant differences were not observed in the expression of miRNA-24-3p, while miRNA-542-3p and 196a-5p were not detected in the sample group. No significant difference was observed in the expression of miRNAs in serum samples. The assessment of the diagnostic potential of microRNAs included ROC curve analysis, which revealed that miR-21-5p had an area under the curve (AUC) of 0.803, while miR-31-5p obtained an AUC of 0.777. The results also identified differential expression among the proteins survivin, PLK1, and p63, all of which showed increased expression in tumour tissue. Additionally, a correlation was observed between the expression of miR-21-5p and the protein p40 (chi-square: p=0.047; Spearman correlation: r=0.402; p=0.023). In conclusion, the results suggest that miR-21-5p and miR-31-5p may be potential diagnostic biomarkers for tongue squamous cell carcinoma, providing a foundation for further exploration for large-scale studies to explore miRNA-protein correlations, considering the site specificity of miRNAs.
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spelling Camillo, Cláudia Malheiros Coutinhohttps://orcid.org/0000-0001-9016-2668http://lattes.cnpq.br/3184503163639480Zeidler, Sandra Lúcia Ventorin vonhttps://orcid.org/0000-0002-8897-5747http://lattes.cnpq.br/5785612863130498Có, Anna Clara Gregóriohttps://orcid.org/0000-0002-7737-0371http://lattes.cnpq.br/3678557620411441Nunes, Fabio Daumashttps://orcid.org/0000-0002-7785-6785http://lattes.cnpq.br/4909755821591847Errera, Flavia Imbroisi Vallehttps://orcid.org/0000-0002-8069-6372http://lattes.cnpq.br/9337327437538048Arantes, Lidia Maria Rebolho Batistahttps://orcid.org/0000-0001-8230-1218http://lattes.cnpq.br/2019308149950531Paula, Flavia dehttps://orcid.org/0000-0001-8679-2982http://lattes.cnpq.br/79132014506636832024-05-29T20:55:26Z2024-05-29T20:55:26Z2023-11-22Oral squamous cell carcinoma (OSCC) is the 16th most commonly diagnosed form of cancer globally, with a higher prevalence in the tongue compared to other areas of the oral cavity. However, the lack of effective biomarkers for diagnosis, especially for precancerous lesions, poses a limitation, as visual or histological examination cannot predict the progression of dysplastic lesions, making it difficult to determine whether they will develop into cancer or return to normal epithelium. In this context, the present research aims to investigate molecular targets that may indicate the irreversible transformation of these cells, to provide a basis for broader studies aimed at using these targets as biomarkers for early OSCC diagnosis. To achieve this goal, this experimental study addressed the evaluation of the expression of a panel of microRNAs and proteins in tumour tissues and adjacent tumour-adjacent epithelium obtained from patients with tongue squamous cell carcinoma and oral epithelium from healthy individuals. Additionally, the research explored the association between these biomarkers, seeking to determine their potential application as diagnostic biomarkers for tongue squamous cell carcinoma. A total of 75 cases of tongue squamous cell carcinoma and adjacent tumouradjacent epithelium were included in the study, and the expression of the proteins survivin, Bcl-2, PLK1, p16, p40, p63, EGFR, and cyclin D1 was analysed by immunohistochemistry. The analysis of the microRNA panel's expression involved 31 samples of tongue squamous cell carcinoma, 10 samples of healthy gingival tissue, and 10 samples of serum from healthy individuals, as well as 7 samples of serum from patients diagnosed with OSCC, using the RT-qPCR technique. In silico analysis by bioinformatics validated the findings related to the expression of differentially expressed microRNAs in the sample group. The results showed differences in the expression of miRNA-31-5p (p<0.001) and miRNA-21-5p (p=0.001) in tumour samples compared to control samples. Significant differences were not observed in the expression of miRNA-24-3p, while miRNA-542-3p and 196a-5p were not detected in the sample group. No significant difference was observed in the expression of miRNAs in serum samples. The assessment of the diagnostic potential of microRNAs included ROC curve analysis, which revealed that miR-21-5p had an area under the curve (AUC) of 0.803, while miR-31-5p obtained an AUC of 0.777. The results also identified differential expression among the proteins survivin, PLK1, and p63, all of which showed increased expression in tumour tissue. Additionally, a correlation was observed between the expression of miR-21-5p and the protein p40 (chi-square: p=0.047; Spearman correlation: r=0.402; p=0.023). In conclusion, the results suggest that miR-21-5p and miR-31-5p may be potential diagnostic biomarkers for tongue squamous cell carcinoma, providing a foundation for further exploration for large-scale studies to explore miRNA-protein correlations, considering the site specificity of miRNAs.O carcinoma epidermoide oral (CEO) é o 16º tipo mais incidente de câncer no mundo, sendo a língua a localização anatômica mais frequentemente afetada. No entanto, a ausência de biomarcadores eficazes para o diagnóstico, especialmente de lesões precursoras, representa uma limitação, uma vez que o exame visual ou histológico não são capazes de predizer a evolução das lesões displasicas, tornando difícil determinar se estas irão progredir para o desenvolvimento do câncer ou retornar a um epitélio normal. Nesse contexto, a presente pesquisa se propõe a investigar alvos moleculares que possam indicar a transformação irreversível dessas células, com o intuito de fornecer a base para estudos mais amplos visando a utilização desses alvos como biomarcadores para o diagnóstico precoce do CEO. Para alcançar esse objetivo, este estudo experimental abordou a avaliação da expressão de um painel de microRNAs e de proteínas em tecidos tumorais e epitélio adjacente ao tumor obtidos de pacientes com carcinoma epidermoide de língua e tecidos de epitélio oral provenientes de indivíduos saudáveis. Além disso, a pesquisa explorou a associação entre esses biomarcadores, buscando determinar sua potencial aplicação como biomarcadores diagnóstico para carcinoma epidermoide de língua. Um total de 75 casos de carcinoma epidermoide de língua e epitélio adjacente ao tumor foram incluídos no estudo, e a expressão das proteínas survivina, Bcl-2, PLK1, p16, p40, p63, EGFR e ciclina D1 foi analisada por imunohistoquímica. A análise da expressão do painel de microRNAs envolveu 31 amostras de carcinoma epidermoide de língua, 10 amostras de tecido gengival saudável e 10 amostras de soro de indivíduos saudáveis, bem como 7 amostras de soro de pacientes diagnosticados com CEO, empregando a técnica de RT-qPCR. A análise in silico por bioinformática validou os achados relativos à expressão dos microRNAs diferencialmente expressos no grupo amostral. Os resultados mostraram diferença na expressão dos miRNA-31-5p (p<0,001) e miRNA-21-5p (p=0,001) nas amostras tumorais em comparação com as amostras controle. Diferença significativa não foi observada na expressão do miRNA-24-3p, enquanto os miRNA-542-3p e 196a-5p não foram detectados no grupo amostral. Não foi observada diferença significante entre a expressão dos miRNAs em amostras de soro. A avaliação do potencial de diagnóstico dos microRNAs incluiu análise da curva ROC, a qual revelou que o miR-21-5p apresentou uma área sob a curva (AUC) de 0,803, enquanto o miR-31-5p obteve uma AUC de 0,777. O resultado também permitiu identificar expressão diferencial entre as proteínas survivina, PLK1 e p63, todas elas com expressão aumentada no tecido tumoral. Além disso, observou-se uma correlação entre a expressão do miR-21-5p e a proteína p40 (qui-quadrado: p=0,047; correlação de Spearman: r=0,402; p=0,023). Em conclusão, os resultados obtidos sugerem que o miR-21-5p e o miR-31-5p podem ser potenciais biomarcadores de diagnóstico para o Carcinoma epidermoide de língua, sendo este o ponto de partida para a elaboração de estudos em larga escala para explorar as correlações miRNA-proteína, considerando a sítio especificidade dos miRNAs.Fundação de Amparo à Pesquisa do Espírito Santo (FAPES)Texthttp://repositorio.ufes.br/handle/10/12554porUniversidade Federal do Espírito SantoDoutorado em BiotecnologiaPrograma de Pós-Graduação em BiotecnologiaUFESBRCentro de Ciências da Saúdeembargoed access info:eu-repo/semantics/openAccessBiotecnologiaMiRNAsMiR-21-5pMiR-31-5pBiomarcadoresCarcinoma epidermoide oralLínguaCâncer da cavidade oralSurvivinaPLK1 e p63Oral squamous cell carcinomaMicroRNAsAvaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de línguaEvaluation of microRNA and protein expression as diagnostic biomarkers in tongue squamous cell carcinomainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFESannaclaragc@gmail.comORIGINALEMBARGADO-RESTRITO.pdfapplication/pdf275372http://repositorio.ufes.br/bitstreams/870e7a07-2980-404f-bb51-a8ff51970679/downloadf19515a01cb1c30076d7f7ba8c48dd73MD51ORIGINALORIGINALORIGINALORIGINALORIGINALORIGINAL10/125542024-09-26 10:45:05.571oai:repositorio.ufes.br:10/12554http://repositorio.ufes.brRepositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-10-15T17:54:19.908033Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)false
dc.title.none.fl_str_mv Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
dc.title.alternative.none.fl_str_mv Evaluation of microRNA and protein expression as diagnostic biomarkers in tongue squamous cell carcinoma
title Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
spellingShingle Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
Có, Anna Clara Gregório
Biotecnologia
MiRNAs
MiR-21-5p
MiR-31-5p
Biomarcadores
Carcinoma epidermoide oral
Língua
Câncer da cavidade oral
Survivina
PLK1 e p63
Oral squamous cell carcinoma
MicroRNAs
title_short Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
title_full Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
title_fullStr Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
title_full_unstemmed Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
title_sort Avaliação da expressão de microRNAs e proteínas como biomarcadores de diagnóstico em carcinoma epidermoide de língua
author Có, Anna Clara Gregório
author_facet Có, Anna Clara Gregório
author_role author
dc.contributor.authorID.none.fl_str_mv https://orcid.org/0000-0002-7737-0371
dc.contributor.authorLattes.none.fl_str_mv http://lattes.cnpq.br/3678557620411441
dc.contributor.advisor-co1.fl_str_mv Camillo, Cláudia Malheiros Coutinho
dc.contributor.advisor-co1ID.fl_str_mv https://orcid.org/0000-0001-9016-2668
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/3184503163639480
dc.contributor.advisor1.fl_str_mv Zeidler, Sandra Lúcia Ventorin von
dc.contributor.advisor1ID.fl_str_mv https://orcid.org/0000-0002-8897-5747
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/5785612863130498
dc.contributor.author.fl_str_mv Có, Anna Clara Gregório
dc.contributor.referee1.fl_str_mv Nunes, Fabio Daumas
dc.contributor.referee1ID.fl_str_mv https://orcid.org/0000-0002-7785-6785
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/4909755821591847
dc.contributor.referee2.fl_str_mv Errera, Flavia Imbroisi Valle
dc.contributor.referee2ID.fl_str_mv https://orcid.org/0000-0002-8069-6372
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/9337327437538048
dc.contributor.referee3.fl_str_mv Arantes, Lidia Maria Rebolho Batista
dc.contributor.referee3ID.fl_str_mv https://orcid.org/0000-0001-8230-1218
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/2019308149950531
dc.contributor.referee4.fl_str_mv Paula, Flavia de
dc.contributor.referee4ID.fl_str_mv https://orcid.org/0000-0001-8679-2982
dc.contributor.referee4Lattes.fl_str_mv http://lattes.cnpq.br/7913201450663683
contributor_str_mv Camillo, Cláudia Malheiros Coutinho
Zeidler, Sandra Lúcia Ventorin von
Nunes, Fabio Daumas
Errera, Flavia Imbroisi Valle
Arantes, Lidia Maria Rebolho Batista
Paula, Flavia de
dc.subject.cnpq.fl_str_mv Biotecnologia
topic Biotecnologia
MiRNAs
MiR-21-5p
MiR-31-5p
Biomarcadores
Carcinoma epidermoide oral
Língua
Câncer da cavidade oral
Survivina
PLK1 e p63
Oral squamous cell carcinoma
MicroRNAs
dc.subject.por.fl_str_mv MiRNAs
MiR-21-5p
MiR-31-5p
Biomarcadores
Carcinoma epidermoide oral
Língua
Câncer da cavidade oral
Survivina
PLK1 e p63
Oral squamous cell carcinoma
MicroRNAs
description Oral squamous cell carcinoma (OSCC) is the 16th most commonly diagnosed form of cancer globally, with a higher prevalence in the tongue compared to other areas of the oral cavity. However, the lack of effective biomarkers for diagnosis, especially for precancerous lesions, poses a limitation, as visual or histological examination cannot predict the progression of dysplastic lesions, making it difficult to determine whether they will develop into cancer or return to normal epithelium. In this context, the present research aims to investigate molecular targets that may indicate the irreversible transformation of these cells, to provide a basis for broader studies aimed at using these targets as biomarkers for early OSCC diagnosis. To achieve this goal, this experimental study addressed the evaluation of the expression of a panel of microRNAs and proteins in tumour tissues and adjacent tumour-adjacent epithelium obtained from patients with tongue squamous cell carcinoma and oral epithelium from healthy individuals. Additionally, the research explored the association between these biomarkers, seeking to determine their potential application as diagnostic biomarkers for tongue squamous cell carcinoma. A total of 75 cases of tongue squamous cell carcinoma and adjacent tumouradjacent epithelium were included in the study, and the expression of the proteins survivin, Bcl-2, PLK1, p16, p40, p63, EGFR, and cyclin D1 was analysed by immunohistochemistry. The analysis of the microRNA panel's expression involved 31 samples of tongue squamous cell carcinoma, 10 samples of healthy gingival tissue, and 10 samples of serum from healthy individuals, as well as 7 samples of serum from patients diagnosed with OSCC, using the RT-qPCR technique. In silico analysis by bioinformatics validated the findings related to the expression of differentially expressed microRNAs in the sample group. The results showed differences in the expression of miRNA-31-5p (p<0.001) and miRNA-21-5p (p=0.001) in tumour samples compared to control samples. Significant differences were not observed in the expression of miRNA-24-3p, while miRNA-542-3p and 196a-5p were not detected in the sample group. No significant difference was observed in the expression of miRNAs in serum samples. The assessment of the diagnostic potential of microRNAs included ROC curve analysis, which revealed that miR-21-5p had an area under the curve (AUC) of 0.803, while miR-31-5p obtained an AUC of 0.777. The results also identified differential expression among the proteins survivin, PLK1, and p63, all of which showed increased expression in tumour tissue. Additionally, a correlation was observed between the expression of miR-21-5p and the protein p40 (chi-square: p=0.047; Spearman correlation: r=0.402; p=0.023). In conclusion, the results suggest that miR-21-5p and miR-31-5p may be potential diagnostic biomarkers for tongue squamous cell carcinoma, providing a foundation for further exploration for large-scale studies to explore miRNA-protein correlations, considering the site specificity of miRNAs.
publishDate 2023
dc.date.issued.fl_str_mv 2023-11-22
dc.date.accessioned.fl_str_mv 2024-05-29T20:55:26Z
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dc.publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Doutorado em Biotecnologia
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dc.publisher.department.fl_str_mv Centro de Ciências da Saúde
publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Doutorado em Biotecnologia
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