Construction of human mucosa oral epithelial cell lines overexpressing telomerase reverse transcriptase gene mediated by lentivirus / 华西口腔医学杂志
West China Journal of Stomatology
;
(6): 443-447, 2016.
Artículo
en Chino
| WPRIM
| ID: wpr-317788
ABSTRACT
<p><b>OBJECTIVE</b>To construct a cell line of oral mucosa epithelial cells that stably express human telomerase reverse transcriptase (hTERT) by lentiviral vectors, approaches for the establishment of stable and efficient immortalized oral mucosa epithelial cell lines were explored.</p><p><b>METHODS</b>Whole RNA was extracted from 293T cells. The hTERT gene was amplified by polymerase chain reaction (PCR) and cloned into the lentiviral vector as pLVX-puro-hTERT. The lentivirus particles were successfully packaged and used to infect primary oral epithelial cells. The positive cell clones were selected by puromycin. Finally, the expression of hTERT was examined by real-time fluorescent quantitative PCR (qRT-PCR) and Western blot analysis.</p><p><b>RESULTS</b>The sequencing results confirmed the construction of the recombinant lentivirus pLVX-puro-hTERT. The morphology of infected cells was similar to that of normal oral mucosal epithelial cells, with a cobble stone-like appearance. The qRT-PCR and Western blot results showed that hTERT was overexpressed in infected cells compared with the normal group (P<0.05).</p><p><b>CONCLUSIONS</b>The oral epithelial cell line with stable expression of hTERT was successfully established by the lentivirus, which provides an experimental basis for the establishment of a highly efficient and stable oral epithelial immortalized cell line.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Lentivirus
/
Telomerasa
/
Células HEK293
/
Reacción en Cadena en Tiempo Real de la Polimerasa
/
Boca
/
Membrana Mucosa
Límite:
Humanos
Idioma:
Chino
Revista:
West China Journal of Stomatology
Año:
2016
Tipo del documento:
Artículo
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