Artigo

Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis

Human periodontal ligament fibroblast (hPLF) cells play an important role in maintaining oral cavity homeostasis with special function in tissue regeneration and maintenance of dental alveoli. Although their primary cell cultures are considered a good experimental model with no genetic changes, the...

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Autor principal: Nogueira, Lygia S
Outros Autores: Vasconcelos, Carolina P, Mitre, Geovanni Pereira, Bittencourt, Leonardo Oliveira, Pla?a, Jessica Rodrigues, Kataoka, Maria Sueli da Silva, Pinheiro, Jo?o de Jesus Viana, Garlet, Gustavo Pompermaier, Oliveira, Edivaldo Herculano Corr?a de, Lima, Rafael R
Grau: Artigo
Idioma: eng
Publicado em: Frontiers Media 2021
Assuntos:
Doen?as Dent?rias
Ligamento Periodontal / citologia
Telomerase
Express?o G?nica
Acesso em linha: http://patua.iec.gov.br//handle/iec/4328
id ir-iec-4328
recordtype dspace
spelling ir-iec-43282021-06-28T12:29:26Z Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis Nogueira, Lygia S Vasconcelos, Carolina P Mitre, Geovanni Pereira Bittencourt, Leonardo Oliveira Pla?a, Jessica Rodrigues Kataoka, Maria Sueli da Silva Pinheiro, Jo?o de Jesus Viana Garlet, Gustavo Pompermaier Oliveira, Edivaldo Herculano Corr?a de Lima, Rafael R Doen?as Dent?rias Ligamento Periodontal / citologia Telomerase Express?o G?nica Human periodontal ligament fibroblast (hPLF) cells play an important role in maintaining oral cavity homeostasis with special function in tissue regeneration and maintenance of dental alveoli. Although their primary cell cultures are considered a good experimental model with no genetic changes, the finite life span may limit some experimental designs. The immortalization process increases cell life span but may cause genetic changes and chromosomal instability, resulting in direct effects on physiological cell responses. In this way, we aimed to investigate the global gene expression of hPLFs after the immortalization process by the ectopic expression of the catalytic subunit of the enzyme telomerase reverse transcriptase (hTERT) through transcriptome analysis. The embryonic origin of the primary culture of hPLF cells and immortalized hPLF-hTERT was also tested by vimentin staining, hTERT synthesis evaluated by indirect immunocytochemistry, analysis of cell proliferation, and morphology. The results indicated that hPLFs and hPLF-hTERT were positive for vimentin. On the 20th cell passage, hPLFs were in senescence, while hPLF-hTERT maintained their proliferation and morphology characteristics. At the same passage, hPLF-hTERT presented a significant increase in hTERT synthesis, but transcriptome did not reveal overexpression of the hTERT gene. Fifty-eight genes had their expression altered (11 upregulated and 47 downregulated) with the absence of changes in the key genes related to these cell types and in the main cancer-associated genes. In addition, the increase in hTERT protein expression without the overexpression of its gene indicates posttranscriptional level regulation. Successful immortalization of hPLFs through the ectopic expression of hTERT encourages further studies to design experimental protocols to investigate clinical questions from a translational perspective. This study was partially supported by a grant provided by CNPq (Brazilian National Council for Scientific and Technological Development, 435093/2018-5) and CAPES/PROCAD - Higher Education Improvement Coordination (23038.005350/2018-78; Finance Code 001). Pr?-Reitoria dePesquisa e P?s-gradua??o from the Federal University of Par? (PROPESP-UFPA). 2021-06-21T13:29:40Z 2021-06-21T13:29:40Z 2021 Artigo NOGUEIRA, Lygia S. et al. Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis. Frontiers in Molecular Biosciences, v. 8, n. 679548, June 2021. 2296-889X http://patua.iec.gov.br//handle/iec/4328 10.3389/fmolb.2021.679548 eng Acesso Aberto application/pdf Frontiers Media
institution Instituto Evandro Chagas (IEC)
collection PATUA
language eng
topic Doen?as Dent?rias
Ligamento Periodontal / citologia
Telomerase
Express?o G?nica
spellingShingle Doen?as Dent?rias
Ligamento Periodontal / citologia
Telomerase
Express?o G?nica
Nogueira, Lygia S
Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis
topic_facet Doen?as Dent?rias
Ligamento Periodontal / citologia
Telomerase
Express?o G?nica
description Human periodontal ligament fibroblast (hPLF) cells play an important role in maintaining oral cavity homeostasis with special function in tissue regeneration and maintenance of dental alveoli. Although their primary cell cultures are considered a good experimental model with no genetic changes, the finite life span may limit some experimental designs. The immortalization process increases cell life span but may cause genetic changes and chromosomal instability, resulting in direct effects on physiological cell responses. In this way, we aimed to investigate the global gene expression of hPLFs after the immortalization process by the ectopic expression of the catalytic subunit of the enzyme telomerase reverse transcriptase (hTERT) through transcriptome analysis. The embryonic origin of the primary culture of hPLF cells and immortalized hPLF-hTERT was also tested by vimentin staining, hTERT synthesis evaluated by indirect immunocytochemistry, analysis of cell proliferation, and morphology. The results indicated that hPLFs and hPLF-hTERT were positive for vimentin. On the 20th cell passage, hPLFs were in senescence, while hPLF-hTERT maintained their proliferation and morphology characteristics. At the same passage, hPLF-hTERT presented a significant increase in hTERT synthesis, but transcriptome did not reveal overexpression of the hTERT gene. Fifty-eight genes had their expression altered (11 upregulated and 47 downregulated) with the absence of changes in the key genes related to these cell types and in the main cancer-associated genes. In addition, the increase in hTERT protein expression without the overexpression of its gene indicates posttranscriptional level regulation. Successful immortalization of hPLFs through the ectopic expression of hTERT encourages further studies to design experimental protocols to investigate clinical questions from a translational perspective.
format Artigo
author Nogueira, Lygia S
author2 Vasconcelos, Carolina P
Mitre, Geovanni Pereira
Bittencourt, Leonardo Oliveira
Pla?a, Jessica Rodrigues
Kataoka, Maria Sueli da Silva
Pinheiro, Jo?o de Jesus Viana
Garlet, Gustavo Pompermaier
Oliveira, Edivaldo Herculano Corr?a de
Lima, Rafael R
author2Str Vasconcelos, Carolina P
Mitre, Geovanni Pereira
Bittencourt, Leonardo Oliveira
Pla?a, Jessica Rodrigues
Kataoka, Maria Sueli da Silva
Pinheiro, Jo?o de Jesus Viana
Garlet, Gustavo Pompermaier
Oliveira, Edivaldo Herculano Corr?a de
Lima, Rafael R
title Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis
title_short Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis
title_full Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis
title_fullStr Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis
title_full_unstemmed Gene expression profile in immortalized human periodontal ligament fibroblasts through hTERT ectopic expression: transcriptome and bioinformatic analysis
title_sort gene expression profile in immortalized human periodontal ligament fibroblasts through htert ectopic expression: transcriptome and bioinformatic analysis
publisher Frontiers Media
publishDate 2021
url http://patua.iec.gov.br//handle/iec/4328
_version_ 1717584457620258816
score 11.755432

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