Construction and selection of siRNA expression cassettes targeting human telomerase reverse transcriptase gene in vitro / 中华口腔医学杂志
Chinese Journal of Stomatology
;
(12): 403-406, 2006.
Artigo
em Chinês
| WPRIM
| ID: wpr-354359
ABSTRACT
<p><b>OBJECTIVE</b>To determine whether the human telomerase reverse transcriptase (hTERT) gene silencing could be effectively induced by PCR-derived siRNA expression cassettes (SEC) transfected by the fifth generation polyamidoamine dendrimer (G5 PAMAM-D) in Tca8113 cells.</p><p><b>METHODS</b>Four SEC were rationally designed and constructed based on a two-step PCR reaction. The SEC were then transferred into Tca8113 cells using G5 PAMAM-D, and hTERT expression was investigated by real-time fluorescence-quantitative reverse transcriptase-PCR and western blot analysis.</p><p><b>RESULTS</b>The RNA interference effects of the SEC targeted for varying hTERT mRNA positions showed a significant disparity. Among them, SEC-A revealed the most potent inhibitory effects (above 95% of reduction), followed by SEC-D and SEC-C, and SEC-B had no effect on hTERT expression (P > 0.05). That the endogenous hTERT gene silencing induced by G5 PAMAM dendrimer-mediated SEC-A was highly sequence-specific, and multiple transfection as well as properties of the vectors were routinely attributable to the specific suppression.</p><p><b>CONCLUSIONS</b>Specific inhibition of endogenous hTERT expression by use of a PCR-based short hairpin siRNA technique and dendrimer transfer system may serve as a novel strategy for treatment of tongue cancers expressing hTERT in vitro.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Neoplasias da Língua
/
Carcinoma de Células Escamosas
/
Transfecção
/
Expressão Gênica
/
Telomerase
/
RNA Interferente Pequeno
/
Linhagem Celular Tumoral
/
Vetores Genéticos
/
Genética
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Stomatology
Ano de publicação:
2006
Tipo de documento:
Artigo
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