Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal

Detalhes bibliográficos
Autor(a) principal: Proença, Marcela Alcântara [UNESP]
Data de Publicação: 2017
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://hdl.handle.net/11449/150007
Resumo: Colorectal cancer (CRC) is associated with pathogens such as Fusobacterium nucleatum (Fn), which can provide a favorable microenvironment for tumorigenesis due to inflammatory changes. In order to understand the effect of Fn on the microenvironment of intestinal lesions, the relative quantification (RQ) of this bacterium was evaluated in samples of colorectal adenoma tissue (CRA) and CCR, as well as its correlation with the mRNA expression of inflammatory mediators (TLR2, TLR4, NFKB1, TNF, IL1B, IL6 e IL8), and microRNAs (miR-21-3p, miR-22-3p, miR-28-5p, miR-34a-5p e miR-135b-5p) involved in the inflammatory response and carcinogenesis. A miRNA: mRNA interaction network was also delineated to aid in the understanding of miRNA participation in the carcinogenic process. DNA and RNA were extracted from 27 fresh tissue samples of CRA and 43 of CRC and their respective adjacent normal ones. Fn and mRNA levels of the inflammatory mediators and miRNAs were quantified by quantitative real-time PCR (qPCR). Elevated levels of Fn were detected in CRA (RQ=5.64 and more markedly in CRC (RQ=8.67). High mRNA expression of TLR4, IL1B, IL8 and miR-135b in CRA, and of TLR2, IL1B, IL6, IL8, miR-34a and miR-135b in CRC were observed in comparison with their respective normal tissues. In addition, miR-22 and miR-28 were found downregulated in CRC. The mRNA expression of IL1B, IL6, IL8 and miR-22 was positively correlated with the quantification of Fn in CRC. The mRNA expression of miR-135b and TNF was inversely correlated in tumor tissue, suggesting TNF as a possible target of this miRNA in CRC. The interaction miRNA:mRNA network suggest that the increased expression of miR-34a in CRC can regulates TLR4 favoring the recognition of Fn by TLR2, which is upregulated in tumor tissue, whereas in CRA this recognition must be mainly via TLR4. These results suggest an increase in Fn colonization during the progression of the adenoma-adenocarcinoma pathway, indicating it as a risk factor for colorectal carcinogenesis. The abundance of this bacterium in the gut promotes increased expression of inflammatory mediators such as IL1B, IL6 and IL8 through their recognition by TLR2 or TLR4, depending on the interaction with differentially expressed miRNAs.
id UNSP_9e295698f06f2ec22a2ec77004ed4203
oai_identifier_str oai:repositorio.unesp.br:11449/150007
network_acronym_str UNSP
network_name_str Repositório Institucional da UNESP
repository_id_str 2946
spelling Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretalEvaluation of the influence of Fusobacterium nucleatum on the modulation of cytokines and microRNAs in adenoma and colorectal cancerCâncer colorretalAdenoma colorretalFusobacterium nucleatumInflamaçãoCitocinasMicroRNAsColorectal cancerColorectal adenomaInflammationCytokinesColorectal cancer (CRC) is associated with pathogens such as Fusobacterium nucleatum (Fn), which can provide a favorable microenvironment for tumorigenesis due to inflammatory changes. In order to understand the effect of Fn on the microenvironment of intestinal lesions, the relative quantification (RQ) of this bacterium was evaluated in samples of colorectal adenoma tissue (CRA) and CCR, as well as its correlation with the mRNA expression of inflammatory mediators (TLR2, TLR4, NFKB1, TNF, IL1B, IL6 e IL8), and microRNAs (miR-21-3p, miR-22-3p, miR-28-5p, miR-34a-5p e miR-135b-5p) involved in the inflammatory response and carcinogenesis. A miRNA: mRNA interaction network was also delineated to aid in the understanding of miRNA participation in the carcinogenic process. DNA and RNA were extracted from 27 fresh tissue samples of CRA and 43 of CRC and their respective adjacent normal ones. Fn and mRNA levels of the inflammatory mediators and miRNAs were quantified by quantitative real-time PCR (qPCR). Elevated levels of Fn were detected in CRA (RQ=5.64 and more markedly in CRC (RQ=8.67). High mRNA expression of TLR4, IL1B, IL8 and miR-135b in CRA, and of TLR2, IL1B, IL6, IL8, miR-34a and miR-135b in CRC were observed in comparison with their respective normal tissues. In addition, miR-22 and miR-28 were found downregulated in CRC. The mRNA expression of IL1B, IL6, IL8 and miR-22 was positively correlated with the quantification of Fn in CRC. The mRNA expression of miR-135b and TNF was inversely correlated in tumor tissue, suggesting TNF as a possible target of this miRNA in CRC. The interaction miRNA:mRNA network suggest that the increased expression of miR-34a in CRC can regulates TLR4 favoring the recognition of Fn by TLR2, which is upregulated in tumor tissue, whereas in CRA this recognition must be mainly via TLR4. These results suggest an increase in Fn colonization during the progression of the adenoma-adenocarcinoma pathway, indicating it as a risk factor for colorectal carcinogenesis. The abundance of this bacterium in the gut promotes increased expression of inflammatory mediators such as IL1B, IL6 and IL8 through their recognition by TLR2 or TLR4, depending on the interaction with differentially expressed miRNAs.O câncer colorretal (CCR) está associado à patógenos como Fusobacterium nucleatum (Fn), que podem proporcionar um microambiente favorável para a tumorigênese em decorrência de alterações inflamatórias. Visando compreender o efeito de Fn no microambiente de lesões intestinais, avaliou-se a quantificação relativa (RQ) dessa bactéria em amostras de tecido de adenoma colorretal (ACR) e CCR, bem como sua correlação com a expressão de RNAm de mediadores inflamatórios (TLR2, TLR4, NFKB1, TNF, IL1B, IL6 e IL8) e de microRNAs (miRNAs) (miR-21-3p, miR-22-3p, miR-28-5p, miR-34a-5p e miR-135b-5p) envolvidos na resposta inflamatória e carcinogênese. Também delineou-se uma rede de interação miRNA:RNAm para auxiliar na compreensão da participação dos miRNAs no processo carcinogênico. Foram extraídos o DNA e o RNA de 27 amostras de tecido fresco de ACR e 43 de CCR e suas respectivas normais adjacentes. Os níveis de DNA de Fn e de RNAm dos mediadores inflamatórios e miRNAs foram quantificados por PCR quantitativo em tempo real (qPCR). Níveis elevados de Fn foram detectados em ACR (RQ=5,64) e mais acentuadamente em CCR (RQ=8,67). Observou-se expressão elevada do RNAm de TLR4, IL1B, IL8 e miR-135b em ACR, e de TLR2, IL1B, IL6, IL8, miR-34a e miR-135b em CCR em comparação com seus respectivos tecidos normais. Além disso, miR-22 e miR-28 foram encontrados com expressão reduzida em CCR. A expressão de RNAm de IL1B, IL6, IL8 e miR-22 foi positivamente correlacionada com a quantificação de Fn em CCR. A expressão de RNAm de miR-135b e TNF foi inversamente correlacionada no tecido tumoral, sugerindo TNF como um possível alvo deste miRNA em CCR. As interações miRNA:RNAm sugerem que a expressão aumentada de miR-34a em CCR regula TLR4 favorecendo o reconhecimento de Fn por TLR2, que encontra-se mais expresso no tecido tumoral, enquanto em ACR este reconhecimento deve ser principalmente via TLR4. Esses resultados sugerem um aumento da colonização de Fn durante a progressão da via adenoma-adenocarcinoma indicando-a como um fator de risco para a carcionogênese colorretal. A abundância dessa bactéria no intestino promove o aumento da expressão de mediadores inflamatórios como IL1B, IL6 e IL8 através do seu reconhecimento por TLR2 ou TLR4, conforme a interação com miRNAs diferentemente expressos.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Estadual Paulista (Unesp)Silva, Ana Elizabete [UNESP]Hughes, David J.Universidade Estadual Paulista (Unesp)Proença, Marcela Alcântara [UNESP]2017-03-29T19:52:57Z2017-03-29T19:52:57Z2017-03-03info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/11449/15000700088316433004153023P5porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESP2023-10-17T06:07:14Zoai:repositorio.unesp.br:11449/150007Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T15:10:36.412716Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
Evaluation of the influence of Fusobacterium nucleatum on the modulation of cytokines and microRNAs in adenoma and colorectal cancer
title Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
spellingShingle Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
Proença, Marcela Alcântara [UNESP]
Câncer colorretal
Adenoma colorretal
Fusobacterium nucleatum
Inflamação
Citocinas
MicroRNAs
Colorectal cancer
Colorectal adenoma
Inflammation
Cytokines
title_short Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
title_full Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
title_fullStr Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
title_full_unstemmed Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
title_sort Avaliação da influência de Fusobacterium nucleatum na modulação de citocinas e microRNAs em adenoma e câncer colorretal
author Proença, Marcela Alcântara [UNESP]
author_facet Proença, Marcela Alcântara [UNESP]
author_role author
dc.contributor.none.fl_str_mv Silva, Ana Elizabete [UNESP]
Hughes, David J.
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Proença, Marcela Alcântara [UNESP]
dc.subject.por.fl_str_mv Câncer colorretal
Adenoma colorretal
Fusobacterium nucleatum
Inflamação
Citocinas
MicroRNAs
Colorectal cancer
Colorectal adenoma
Inflammation
Cytokines
topic Câncer colorretal
Adenoma colorretal
Fusobacterium nucleatum
Inflamação
Citocinas
MicroRNAs
Colorectal cancer
Colorectal adenoma
Inflammation
Cytokines
description Colorectal cancer (CRC) is associated with pathogens such as Fusobacterium nucleatum (Fn), which can provide a favorable microenvironment for tumorigenesis due to inflammatory changes. In order to understand the effect of Fn on the microenvironment of intestinal lesions, the relative quantification (RQ) of this bacterium was evaluated in samples of colorectal adenoma tissue (CRA) and CCR, as well as its correlation with the mRNA expression of inflammatory mediators (TLR2, TLR4, NFKB1, TNF, IL1B, IL6 e IL8), and microRNAs (miR-21-3p, miR-22-3p, miR-28-5p, miR-34a-5p e miR-135b-5p) involved in the inflammatory response and carcinogenesis. A miRNA: mRNA interaction network was also delineated to aid in the understanding of miRNA participation in the carcinogenic process. DNA and RNA were extracted from 27 fresh tissue samples of CRA and 43 of CRC and their respective adjacent normal ones. Fn and mRNA levels of the inflammatory mediators and miRNAs were quantified by quantitative real-time PCR (qPCR). Elevated levels of Fn were detected in CRA (RQ=5.64 and more markedly in CRC (RQ=8.67). High mRNA expression of TLR4, IL1B, IL8 and miR-135b in CRA, and of TLR2, IL1B, IL6, IL8, miR-34a and miR-135b in CRC were observed in comparison with their respective normal tissues. In addition, miR-22 and miR-28 were found downregulated in CRC. The mRNA expression of IL1B, IL6, IL8 and miR-22 was positively correlated with the quantification of Fn in CRC. The mRNA expression of miR-135b and TNF was inversely correlated in tumor tissue, suggesting TNF as a possible target of this miRNA in CRC. The interaction miRNA:mRNA network suggest that the increased expression of miR-34a in CRC can regulates TLR4 favoring the recognition of Fn by TLR2, which is upregulated in tumor tissue, whereas in CRA this recognition must be mainly via TLR4. These results suggest an increase in Fn colonization during the progression of the adenoma-adenocarcinoma pathway, indicating it as a risk factor for colorectal carcinogenesis. The abundance of this bacterium in the gut promotes increased expression of inflammatory mediators such as IL1B, IL6 and IL8 through their recognition by TLR2 or TLR4, depending on the interaction with differentially expressed miRNAs.
publishDate 2017
dc.date.none.fl_str_mv 2017-03-29T19:52:57Z
2017-03-29T19:52:57Z
2017-03-03
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/11449/150007
000883164
33004153023P5
url http://hdl.handle.net/11449/150007
identifier_str_mv 000883164
33004153023P5
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
publisher.none.fl_str_mv Universidade Estadual Paulista (Unesp)
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1808128474499514368