New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression

Detalhes bibliográficos
Autor(a) principal: Saraiva-Pava, Kathy D.
Data de Publicação: 2015
Outros Autores: Navabi, Nazanin, Skoog, Emma C, Linden,  Sara K., Oleastro, Mónica, Roxo-Rosa, Mónica
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/23440
Resumo: Supported by Grants from the Fundacao para a Ciencia e a Tecnologia (FCT, Portugal), No. PPCDT/SAL-IMI/57297/2004 and No. PTDC/BIM-MEC/1051/2012; The Swedish Cancer foundation; The Swedish Research Council, No. K2010-79X-21372-01-3; Forska utan djurforsok, Animal Free Research; and by Research fellowship 2011 from the Sociedade Portuguesa de Gastrenterologia (Portugal).
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spelling New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expressionPathogenesisBARRIERESTABLISHMENTHELICOBACTER-PYLORI INFECTIONIN-VITRO MODELDISEASEE-CADHERINPATHOGENESISHelicobacter pylori infectionCANCER CELL-LINESHuman gastric epitheliumNCI-N87 cellsIMMORTALIZATIONCellular modelMUCINHelicobacter pylori infectionPathogenesisHuman gastric epitheliumCellular modelNCI-N87 cellsSDG 3 - Good Health and Well-beingSupported by Grants from the Fundacao para a Ciencia e a Tecnologia (FCT, Portugal), No. PPCDT/SAL-IMI/57297/2004 and No. PTDC/BIM-MEC/1051/2012; The Swedish Cancer foundation; The Swedish Research Council, No. K2010-79X-21372-01-3; Forska utan djurforsok, Animal Free Research; and by Research fellowship 2011 from the Sociedade Portuguesa de Gastrenterologia (Portugal).AIM: To establish a cellular model correctly mimicking the gastric epithelium to overcome the limitation in the study of Helicobacter pylori (H. pylori) infection. METHODS: Aiming to overcome this limitation, clones of the heterogenic cancer-derived NCI-N87 cell line were isolated, by stably-transducing it with the human telomerase reverse-transcriptase (hTERT) catalytic subunit gene. The clones were first characterized regarding their cell growth pattern and phenotype. For that we measured the clones' adherence properties, expression of cell-cell junctions' markers (ZO-1 and E-cadherin) and ability to generate a sustained transepithelial electrical resistance. The gastric properties of the clones, concerning expression of mucins, zymogens and glycan contents, were then evaluated by haematoxylin and eosin staining, Periodic acid Schiff (PAS) and PAS/Alcian Blue-staining, immunocytochemistry and Western blot. In addition, we assessed the usefulness of the hTERT-expressing gastric cell line for H. pylori research, by performing co-culture assays and measuring the IL-8 secretion, by ELISA, upon infection with two H. pylori strains differing in virulence. RESULTS: Compared with the parental cell line, the most promising NCI-hTERT-derived clones (CL5 and CL6) were composed of cells with homogenous phenotype, presented higher relative telomerase activities, better adhesion properties, ability to be maintained in culture for longer periods after confluency, and were more efficient in PAS-reactive mucins secretion. Both clones were shown to produce high amounts of MUC1, MUC2 and MUC13. NCI-hTERT-CL5 mucins were shown to be decorated with blood group H type 2 (BG-H), Lewis-x (Le(x)), Le(y) and Le(a) and, in a less extent, with BG-A antigens, but the former two antigens were not detected in the NCI-hTERT-CL6. None of the clones exhibited detectable levels of MUC6 nor sialylated Le(x) and Le(a) glycans. Entailing good gastric properties, both NCI-hTERT-clones were found to produce pepsinogen-5 and human gastric lipase. The progenitor-like phenotype of NCI-hTERT-CL6 cells was highlighted by large nuclei and by the apical vesicular-like distribution of mucin 5AC and Pg5, supporting the accumulation of mucus-secreting and zymogens-chief mature cells functions. CONCLUSION: These traits, in addition to resistance to microaerobic conditions and good responsiveness to H. pylori co-culture, in a strain virulence-dependent manner, make the NCI-hTERT-CL6 a promising model for future in vitro studies.Centro de Estudos de Doenças Crónicas (CEDOC)NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)RUNSaraiva-Pava, Kathy D.Navabi, NazaninSkoog, Emma CLinden,  Sara K.Oleastro, MónicaRoxo-Rosa, Mónica2017-09-19T22:01:52Z2015-06-072015-06-07T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article18application/pdfhttp://hdl.handle.net/10362/23440eng1007-9327PURE: 486449https://doi.org/10.3748/wjg.v21.i21.6526info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T04:11:42Zoai:run.unl.pt:10362/23440Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:27:47.893729Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
title New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
spellingShingle New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
Saraiva-Pava, Kathy D.
Pathogenesis
BARRIER
ESTABLISHMENT
HELICOBACTER-PYLORI INFECTION
IN-VITRO MODEL
DISEASE
E-CADHERIN
PATHOGENESIS
Helicobacter pylori infection
CANCER CELL-LINES
Human gastric epithelium
NCI-N87 cells
IMMORTALIZATION
Cellular model
MUCIN
Helicobacter pylori infection
Pathogenesis
Human gastric epithelium
Cellular model
NCI-N87 cells
SDG 3 - Good Health and Well-being
title_short New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
title_full New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
title_fullStr New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
title_full_unstemmed New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
title_sort New NCI-N87-derived human gastric epithelial line after human telomerase catalytic subunit over-expression
author Saraiva-Pava, Kathy D.
author_facet Saraiva-Pava, Kathy D.
Navabi, Nazanin
Skoog, Emma C
Linden,  Sara K.
Oleastro, Mónica
Roxo-Rosa, Mónica
author_role author
author2 Navabi, Nazanin
Skoog, Emma C
Linden,  Sara K.
Oleastro, Mónica
Roxo-Rosa, Mónica
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Centro de Estudos de Doenças Crónicas (CEDOC)
NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)
RUN
dc.contributor.author.fl_str_mv Saraiva-Pava, Kathy D.
Navabi, Nazanin
Skoog, Emma C
Linden,  Sara K.
Oleastro, Mónica
Roxo-Rosa, Mónica
dc.subject.por.fl_str_mv Pathogenesis
BARRIER
ESTABLISHMENT
HELICOBACTER-PYLORI INFECTION
IN-VITRO MODEL
DISEASE
E-CADHERIN
PATHOGENESIS
Helicobacter pylori infection
CANCER CELL-LINES
Human gastric epithelium
NCI-N87 cells
IMMORTALIZATION
Cellular model
MUCIN
Helicobacter pylori infection
Pathogenesis
Human gastric epithelium
Cellular model
NCI-N87 cells
SDG 3 - Good Health and Well-being
topic Pathogenesis
BARRIER
ESTABLISHMENT
HELICOBACTER-PYLORI INFECTION
IN-VITRO MODEL
DISEASE
E-CADHERIN
PATHOGENESIS
Helicobacter pylori infection
CANCER CELL-LINES
Human gastric epithelium
NCI-N87 cells
IMMORTALIZATION
Cellular model
MUCIN
Helicobacter pylori infection
Pathogenesis
Human gastric epithelium
Cellular model
NCI-N87 cells
SDG 3 - Good Health and Well-being
description Supported by Grants from the Fundacao para a Ciencia e a Tecnologia (FCT, Portugal), No. PPCDT/SAL-IMI/57297/2004 and No. PTDC/BIM-MEC/1051/2012; The Swedish Cancer foundation; The Swedish Research Council, No. K2010-79X-21372-01-3; Forska utan djurforsok, Animal Free Research; and by Research fellowship 2011 from the Sociedade Portuguesa de Gastrenterologia (Portugal).
publishDate 2015
dc.date.none.fl_str_mv 2015-06-07
2015-06-07T00:00:00Z
2017-09-19T22:01:52Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/23440
url http://hdl.handle.net/10362/23440
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1007-9327
PURE: 486449
https://doi.org/10.3748/wjg.v21.i21.6526
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 18
application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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