Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK
Autor(a) principal: | |
---|---|
Data de Publicação: | 2017 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) |
Texto Completo: | http://repositorio.ufes.br/handle/10/7109 |
Resumo: | Oral cavity squamous cell carcinoma (oral SCC) is the most common type of malignancy that affects an oral cavity, usually driven by the pre-malignant lesions, such as leukoplakia. Smoking is one of the main risk factors for oral SCC. Nicotine is the major natural compound present in tobacco, studies have pointed out that its binding to Nicotinic Acetylcholine Receptors (nAChRs) leads to an increase in the production of Reactive Oxygen Species (ROS) and of growth factors that bind to Receptor Tyrosine Kinase (RTK) triggering the activation of MAPKs and Phosphatidylinositol-3-kinase (PI3K / Akt), culminating in the activation of the HIF-1α protein and CA-IX expression, leading to increased cell proliferation, migration, metastasis and inhibition of tumor cell apoptosis. For this purpose we evaluated the expression of genes ERK1 / 2, PI3K, AKT, HIF-1α and CA-IX in cell culture and SCC9 DOK exposed to different concentrations of nicotine and hypoxia chamber at various times. The effect of increasing concentrations of nicotine and exposure to hypoxia chamber in cell lines of DOK and SCC9, cell viability and expression of the genes in question were performed using the MTS technique and Real Time. The results showed higher expression of HIF-1α, PI3K, AKT, ERK1 / 2 and CA-IX in SCC9 at 24 hours in cultivars with 0.1 mM nicotine concentration when compared to control. Comparing the expression of nicotine concentrations with cultured cells in a hypoxia chamber, a greater 24 h time expression in 0.1 mM of HIF-1α, PI3K, AKT and ERK1 / 2 genes was noted, whereas CA-IX was a low expression of hypoxia chamber. The DOK results show that nicotine causes a small increase over control in the expression of HIF-1a in 5 mM, AKT in 2.5 mM and ERK1 in 0.1 mM in the time of 8 hours, during the 24 hours observed a small increase in ERK2 at 2 mM concentration. Compared hypoxia chamber with the control other concentrations and nicotine, it was observed that only in HIF-1α the expression was lower in the time of 8 hours, but in 24 hours this expression exceeded the expression in the control and in the nicotine exposed cells. It is concluded that nicotine is able to modulate a higher expression of genes in question SCC9 than DOK. |
id |
UFES_ca63cc0a2d50fe0e248a664a48f2ef7e |
---|---|
oai_identifier_str |
oai:repositorio.ufes.br:10/7109 |
network_acronym_str |
UFES |
network_name_str |
Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) |
repository_id_str |
2108 |
spelling |
Silva, Adriana Madeira Alvares daZanini, Surama FreitasSantos, Joaquim Gasparini dosNunes, Fábio DaumasTrivilin, Leonardo Oliveira2018-08-01T21:35:02Z2018-08-012018-08-01T21:35:02Z2017-02-23Oral cavity squamous cell carcinoma (oral SCC) is the most common type of malignancy that affects an oral cavity, usually driven by the pre-malignant lesions, such as leukoplakia. Smoking is one of the main risk factors for oral SCC. Nicotine is the major natural compound present in tobacco, studies have pointed out that its binding to Nicotinic Acetylcholine Receptors (nAChRs) leads to an increase in the production of Reactive Oxygen Species (ROS) and of growth factors that bind to Receptor Tyrosine Kinase (RTK) triggering the activation of MAPKs and Phosphatidylinositol-3-kinase (PI3K / Akt), culminating in the activation of the HIF-1α protein and CA-IX expression, leading to increased cell proliferation, migration, metastasis and inhibition of tumor cell apoptosis. For this purpose we evaluated the expression of genes ERK1 / 2, PI3K, AKT, HIF-1α and CA-IX in cell culture and SCC9 DOK exposed to different concentrations of nicotine and hypoxia chamber at various times. The effect of increasing concentrations of nicotine and exposure to hypoxia chamber in cell lines of DOK and SCC9, cell viability and expression of the genes in question were performed using the MTS technique and Real Time. The results showed higher expression of HIF-1α, PI3K, AKT, ERK1 / 2 and CA-IX in SCC9 at 24 hours in cultivars with 0.1 mM nicotine concentration when compared to control. Comparing the expression of nicotine concentrations with cultured cells in a hypoxia chamber, a greater 24 h time expression in 0.1 mM of HIF-1α, PI3K, AKT and ERK1 / 2 genes was noted, whereas CA-IX was a low expression of hypoxia chamber. The DOK results show that nicotine causes a small increase over control in the expression of HIF-1a in 5 mM, AKT in 2.5 mM and ERK1 in 0.1 mM in the time of 8 hours, during the 24 hours observed a small increase in ERK2 at 2 mM concentration. Compared hypoxia chamber with the control other concentrations and nicotine, it was observed that only in HIF-1α the expression was lower in the time of 8 hours, but in 24 hours this expression exceeded the expression in the control and in the nicotine exposed cells. It is concluded that nicotine is able to modulate a higher expression of genes in question SCC9 than DOK.O carcinoma epidermoide de cavidade oral (CEC oral) é o tipo mais comum de neoplasia maligna que acomete a cavidade oral, normalmente conduzido pelo surgimento de lesões pré-malignas, como a leucoplasia. O uso de tabaco compreende um dos maiores fatores de risco para o seguimento de CEC oral. A nicotina é o maior composto natural presente no tabaco, estudos vem apontando que sua ligação a Receptores Nicotínico de Acetilcolina (nAChRs) leva a um aumento da produção de Espécies Reativas de Oxigênio (ROS) e de fatores de crescimento que se ligam a Receptores Tirosina Quinase (RTK) desencadeando ativação de vias de MAPK’s e fosfatidilinositol-3 quinase (PI3K/Akt), que culminam na ativação da proteína HIF-1α e expressão de CA-IX, conduzindo a um aumento na proliferação celular, migração, metástases e a inibição da apoptose de células tumorais. Com este fim avaliamos a expressão dos genes ERK1/2, PI3K, AKT, HIF1α e CA-IX em cultura celular de SCC9 e DOK expostos a diferentes concentrações de nicotina e a câmara de hipóxia em tempos variados. As análises do efeito das concentrações crescentes de nicotina e a exposição à câmara de hipóxia, nas linhagens celulares de SCC9 e DOK, na viabilidade celular e na expressão dos genes em questão foram realizados através de técnica de MTS e Real time. Os resultados apontam maiores expressões em HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em SCC9 no tempo de 24 horas, em cultivados em concentração de 0,1mM de nicotina, quando comparado com o controle. Comparando a expressão das concentrações de nicotina com células cultivas em câmara de hipóxia, nota-se um maior expressão no tempo de 24 horas em 0,1mM dos genes HIF-1α, PI3K, AKT e ERK1/2, já CA-IX ficou com uma expressão baixo de câmara de hipóxia. Os resultados de DOK demostram que a nicotina causa um pequeno aumento em relação ao controle na expressão de HIF-1α em 5mM, AKT em 2,5mM e ERK1 em 0,1mM no tempo de 8 horas, já para o tempo de 24 horas observou-se um pequeno aumento em ERK2 na concentração de 2mM. Comparado câmara de hipóxia com o controle as demais concentrações e nicotina, observou-se que apenas em HIF-1α a expressão foi menor no tempo de 8 horas, mas em 24 horas essa expressão superou as expressões no controle e nas células expostas a nicotina. Conclui-se que a nicotina é capaz de modular uma maior expressão em SCC9 dos genes em questão do que em DOK.Texthttp://repositorio.ufes.br/handle/10/7109porUniversidade Federal do Espírito SantoMestrado em BiotecnologiaPrograma de Pós-Graduação em BiotecnologiaUFESBRCentro de Ciências da SaúdeOral cavityNicotineCavidade oralNicotinaPI3K/AKTERK1/2HIF-1α. CAIXBiotecnologia61Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOKinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFESORIGINALtese_11105_Dissertação_Joaquim Gasparini dos Santos20170608-144422.pdfapplication/pdf1706051http://repositorio.ufes.br/bitstreams/1c0aeef7-b846-4540-a9a2-6421a1f8a4fa/download2bdf20fca97880e12ac7a7ad4a115c94MD5110/71092024-06-27 10:59:26.63oai:repositorio.ufes.br:10/7109http://repositorio.ufes.brRepositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-06-27T10:59:26Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)false |
dc.title.none.fl_str_mv |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK |
title |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK |
spellingShingle |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK Santos, Joaquim Gasparini dos Oral cavity Nicotine Cavidade oral Nicotina PI3K/AKT ERK1/2 HIF-1α. CAIX Biotecnologia 61 |
title_short |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK |
title_full |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK |
title_fullStr |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK |
title_full_unstemmed |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK |
title_sort |
Influência da nicotina na expressão gênica de HIF-1α, PI3K, AKT, ERK1/2 e CA-IX em linhagens celulares SCC9 e DOK |
author |
Santos, Joaquim Gasparini dos |
author_facet |
Santos, Joaquim Gasparini dos |
author_role |
author |
dc.contributor.advisor-co1.fl_str_mv |
Silva, Adriana Madeira Alvares da |
dc.contributor.advisor1.fl_str_mv |
Zanini, Surama Freitas |
dc.contributor.author.fl_str_mv |
Santos, Joaquim Gasparini dos |
dc.contributor.referee1.fl_str_mv |
Nunes, Fábio Daumas |
dc.contributor.referee2.fl_str_mv |
Trivilin, Leonardo Oliveira |
contributor_str_mv |
Silva, Adriana Madeira Alvares da Zanini, Surama Freitas Nunes, Fábio Daumas Trivilin, Leonardo Oliveira |
dc.subject.eng.fl_str_mv |
Oral cavity Nicotine |
topic |
Oral cavity Nicotine Cavidade oral Nicotina PI3K/AKT ERK1/2 HIF-1α. CAIX Biotecnologia 61 |
dc.subject.por.fl_str_mv |
Cavidade oral Nicotina PI3K/AKT ERK1/2 HIF-1α. CAIX |
dc.subject.cnpq.fl_str_mv |
Biotecnologia |
dc.subject.udc.none.fl_str_mv |
61 |
description |
Oral cavity squamous cell carcinoma (oral SCC) is the most common type of malignancy that affects an oral cavity, usually driven by the pre-malignant lesions, such as leukoplakia. Smoking is one of the main risk factors for oral SCC. Nicotine is the major natural compound present in tobacco, studies have pointed out that its binding to Nicotinic Acetylcholine Receptors (nAChRs) leads to an increase in the production of Reactive Oxygen Species (ROS) and of growth factors that bind to Receptor Tyrosine Kinase (RTK) triggering the activation of MAPKs and Phosphatidylinositol-3-kinase (PI3K / Akt), culminating in the activation of the HIF-1α protein and CA-IX expression, leading to increased cell proliferation, migration, metastasis and inhibition of tumor cell apoptosis. For this purpose we evaluated the expression of genes ERK1 / 2, PI3K, AKT, HIF-1α and CA-IX in cell culture and SCC9 DOK exposed to different concentrations of nicotine and hypoxia chamber at various times. The effect of increasing concentrations of nicotine and exposure to hypoxia chamber in cell lines of DOK and SCC9, cell viability and expression of the genes in question were performed using the MTS technique and Real Time. The results showed higher expression of HIF-1α, PI3K, AKT, ERK1 / 2 and CA-IX in SCC9 at 24 hours in cultivars with 0.1 mM nicotine concentration when compared to control. Comparing the expression of nicotine concentrations with cultured cells in a hypoxia chamber, a greater 24 h time expression in 0.1 mM of HIF-1α, PI3K, AKT and ERK1 / 2 genes was noted, whereas CA-IX was a low expression of hypoxia chamber. The DOK results show that nicotine causes a small increase over control in the expression of HIF-1a in 5 mM, AKT in 2.5 mM and ERK1 in 0.1 mM in the time of 8 hours, during the 24 hours observed a small increase in ERK2 at 2 mM concentration. Compared hypoxia chamber with the control other concentrations and nicotine, it was observed that only in HIF-1α the expression was lower in the time of 8 hours, but in 24 hours this expression exceeded the expression in the control and in the nicotine exposed cells. It is concluded that nicotine is able to modulate a higher expression of genes in question SCC9 than DOK. |
publishDate |
2017 |
dc.date.issued.fl_str_mv |
2017-02-23 |
dc.date.accessioned.fl_str_mv |
2018-08-01T21:35:02Z |
dc.date.available.fl_str_mv |
2018-08-01 2018-08-01T21:35:02Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufes.br/handle/10/7109 |
url |
http://repositorio.ufes.br/handle/10/7109 |
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 |
Text |
dc.publisher.none.fl_str_mv |
Universidade Federal do Espírito Santo Mestrado em Biotecnologia |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Biotecnologia |
dc.publisher.initials.fl_str_mv |
UFES |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Centro de Ciências da Saúde |
publisher.none.fl_str_mv |
Universidade Federal do Espírito Santo Mestrado em Biotecnologia |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) instname:Universidade Federal do Espírito Santo (UFES) instacron:UFES |
instname_str |
Universidade Federal do Espírito Santo (UFES) |
instacron_str |
UFES |
institution |
UFES |
reponame_str |
Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) |
collection |
Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) |
bitstream.url.fl_str_mv |
http://repositorio.ufes.br/bitstreams/1c0aeef7-b846-4540-a9a2-6421a1f8a4fa/download |
bitstream.checksum.fl_str_mv |
2bdf20fca97880e12ac7a7ad4a115c94 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 |
repository.name.fl_str_mv |
Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES) |
repository.mail.fl_str_mv |
|
_version_ |
1804309172523630592 |