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Doctoral Thesis
DOI
https://doi.org/10.11606/T.5.2018.tde-07112018-133022
Document
Author
Full name
Tabatha Gutierrez Prieto Martins Rocha
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2018
Supervisor
Committee
Capelozzi, Vera Luiza (President)
Nagai, Maria Aparecida
Begnami, Maria Dirlei Ferreira de Souza
Haddad, Fábio José
Zerbini, Maria Claudia Nogueira
Title in Portuguese
Avaliação dos genes envolvidos na transição epitelial-mesenquimal e sua relação com a progressão e a invasão tumoral em câncer de pulmão
Keywords in Portuguese
Biomarcador molecular
Câncer de pulmão
Metástase
Prognóstico
Progressão tumoral
Transição epitelial-mesenquimal
Abstract in Portuguese
INTRODUÇÃO: As recidivas e as metástases são os fatores responsáveis pela morte de pacientes com câncer de pulmão (representando 90% dos casos), indicando a necessidade de conhecer as múltiplas vias sinalizadoras envolvidas em sua patogênese, progressão, invasão e metástase. Adenocarcinomas e carcinomas de grandes células invadem primariamente vasos sanguíneos e linfáticos, metastatizando à distância. Já os carcinomas de células escamosas envolvem diretamente o mediastino, sobretudo, linfonodos e pericárdio. Carcinomas neuroendócrinos de baixo grau (carcinoides típicos e atípicos) invadem e metastatizam apesar de serem indolentes, enquanto que carcinomas neuroendócrinos de alto grau (carcinoma neuroendócrino de grandes células e carcinoma de pequenas células), rapidamente metastatizam à distância. Esse espectro diferencial de invasividade nos carcinomas de pulmão ainda não pôde ser definitivamente demarcado por biomarcadores, sobretudo nas variantes neuroendócrinas. Neste aspecto, acredita-se que a EMT esteja mais intimamente envolvida no processo de invasão, despontando como potencial biomarcador. OBJETIVO: Realizar o mapeamento genético da EMT nos carcinomas pulmonares, estabelecendo uma assinatura gênica capaz de prever o potencial de invasão nos diferentes tipos histológicos do câncer de pulmão. MÉTODOS: Espécimes de tumores ressecados cirurgicamente e criopreservados de 60 pacientes foram submetidos a análise de expressão gênica através da qPCR em tempo real, por meio do kit RT2 Profiler PCR array para a EMT com 84 genes de interesse. RESULTADOS: Observou-se a elevada expressão de 24 genes (AKT1, COL1A2, COL3A1, COL5A2, DSP, EGFR, FR11, GSK3B, ILK, ITGA5, ITGAV, ITGB1, JAG1, MAP1B, MMP2, MMP3, SNAI2, SPARC, SPP1, STAT3, TCF3, TGF?3, VPS13A, WNT5A) que participam do processo da EMT e atuam no desenvolvimento da neoplasia pulmonar. Tumores em estádio avançado e com metástases linfonodais apresentaram significante maior expressão dos seguintes genes, COL1A2, COL5A2, DSP, EGFR, FR11, GSK3B, ILK, ITGAV, ITGB1, MAP1B, SNAI2 e VPS13A. Comparando-se carcinomas nãoneuroendócrinos (CEC e AD) e neuroendócrinos (CT, CA, CPPC, CNEGC), observou-se que os neuroendócrinos apresentaram elevada expressão significativa dos genes: FR11, GSK3B, ILK, ITGB1, JAG1, MAP1B e VPS13A. Em relação à sobrevida, a análise univariada demonstrou menor sobrevida para pacientes em estádio avançado do tumor com metástase linfonodal e portador de carcinoma neuroendócrino que apresenta a hiperxpressão de todos os 24 genes da EMT, com exceção dos genes COL3A1 e SPP1. Pela análise multivariada demonstrou-se que pacientes com carcinomas neuroendócrinos com expressão elevada de MMP2 e SPARC apresentaram maior risco de morte (OR 5,41 e 4,94, respectivamente), enquanto que pacientes portadores de carcinomas de células escamosas ou adenocarcinomas com baixa expressão gênica de ILK, SPP1, COL1A2, ITGB1 apresentaram menor risco de morte (OR -7,02; -0,4, -1,3 e -3,02). CONCLUSÃO: Através do presente estudo estabeleceu-se uma assinatura gênica de 24 genes capaz de prever o potencial de agressividade histológica, de invasão e de metástases nos carcinomas pulmonares neuroendócrinos e não-neuroendócrinos
Title in English
Evaluation of genes involved in the epithelial-mesenchymal transition and its relation to tumor progression and invasion in lung cancer
Keywords in English
Epithelial-mesenchymal transition
Lung cancer
Metastasis
Molecular biomarker
Prognosis
Tumor progression
Abstract in English
INTRODUCTION: Metastasis are responsible for the death of 90% of patients with lung cancer, indicating the need to know the multiple signaling pathways involved. Adenocarcinomas (Adc) and large cell carcinomas primarily invade blood vessels with distant metastasis, whereas squamous cell carcinoma (SqCC) involves the mediastinal lymph nodes. Neuroendocrine carcinomas of low-grade (typical and atypical carcinoid) are indolent, while high-grade neuroendocrine carcinoma (large cell neuroendocrine carcinoma and small cell carcinoma) metastasize rapidly. Biomarkers of aggressiveness in lung carcinomas remain to be determined, especially in neuroendocrine variants. In these fields, Epithelial to mesenchymal transition (EMT) genes profile emerge promise as an indicator of invasion and metastasis. AIMS: Carry out the genetic mapping of EMT in lung cancer, establishing a gene signature capable of predicting the invasion potential of different histological types of lung cancer. METHODS: Were included specimens of 60 patients and the EMT gene expression was quantified with a quantitative real-time (RT)- PCR carried out on StepOnePlus(TM) Real-Time PCR System (Applied Biosystems) with RT2 Profiler PCR Array System for the EMT pathway wih 84 target genes. (Qiagen, Dusseldorf, Germany). Associations of the gene signature and clinicopathological features, as well as prognostic factors were evaluated. RESULTS: Was observed high expression of 24 genes (AKT1, COL1A2, COL3A1, COL5A2, DSP, EGFR, FR11, GSK3B, ILK, ITGA5, ITGAV, ITGB1, JAG1, MAP1B, MMP2, MMP3, SNAI2, SPARC, SPP1, STAT3, TCF3, TGFbeta3, VPS13A, WNT5A) that are involved in the EMT process and act in the development of pulmonary neoplasia. Tumors at the advanced stage and with lymph node metastasis shown a significant higher expression of the genes COL1A2, COL5A2, DSP, EGFR, FR11, GSK3B, ILK, ITGAV, ITGB1, MAP1B, SNAI2 and VPS13A. Comparing the histological subtypes non-neuroendocrine (SqCC and AD) and neuroendocrine carcinomas (TC, AT, SCLC, LCNEC), was observed that the neuroendocrine variant shown a higher significant expression of the genes FR11, GSK3B, ILK, ITGB1, JAG1, MAP1B and VPS13A. Regarding survival, univariate analysis showed lower survival for patients in the advanced tumor stage, with lymph node metastasis and with neuroendocrine carcinoma histology that presenting overexpression for all 24 EMT genes, with the exception of the COL3A1 and SPP1 genes. Cox regression controlled by, histological types and gene expression of ILK, MMP2, SPP1, SPARC, COL1A2 and ITGB1 showed that patients with neuroendocrine carcinomas with higher expression of MMP2 and SPARC had a higher risk of death (OR 5.41 and 4.94, respectively), whereas patients with squamous cell carcinomas or adenocarcinomas with lower gene expression of ILK, SPP1, COL1A2, ITGB1 showed lower risk of death (OR -7.02, -0.4, -1.3 and -3.02). CONCLUSION: Was established a potential gene signature of 24 genes involved in the EMT process and capable of predicting the histological aggressiveness, invasion and metastasis in neuroendocrine and non-neuroendocrine variants of lung carcinomas
 
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Publishing Date
2018-11-08
 
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