Construction of targeting-Skp2 shRNA plasmids and observation of their inhibitory effect on Tca8113 cells / 中华口腔医学杂志
Chinese Journal of Stomatology
;
(12): 624-628, 2007.
Artigo
em Chinês
| WPRIM
| ID: wpr-359675
ABSTRACT
<p><b>OBJECTIVE</b>To construct the recombinant plasmids expressing Skp2 short hairpin RNA (shRNA) by pRNAT-U6.1/Neo plasmid vector and observe the effects of RNAi-mediated Skp2 gene silencing on Tca8113 cells.</p><p><b>METHODS</b>Five recombinant eukaryotic expression vectors were successfully constructed using pRNAT-U6.1/Neo plasmid vector separately. After they were transfected into Tca8113 cells with PEI, the interference effects no Skp2 and p27 were detected by RT-PCR and Western blot. The cell cycle of Tca8113 cells were tested by flow cytometry. The proliferation of Tca8113 cells were examined by MTT.</p><p><b>RESULTS</b>In Skp2shRNA-2 and Skp2shRNA-3 vectors, the expression of Skp2 protein of Tca8113 cells was down-regulated and p27 protein up-regulated (P < 0.01). The cell number during G1/G 0 phases increased 22% (P < 0.01) and during G(2)/M and S phases the number decreased 10% and 12% (P < 0.01). The proliferation of Tca8113 cells slowed down and the cells number decreased (P < 0.01).</p><p><b>CONCLUSIONS</b>Skp2shRNA-2 and Skp2shRNA-3 vectors of shRNA for Skp2 were successfully constructed. They could influence expression of Skp2 and p27 gene. Skp2 may be a promising target of gene therapy on human tongue squamous cell carcinoma.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Patologia
/
Plasmídeos
/
Neoplasias da Língua
/
Carcinoma de Células Escamosas
/
Transfecção
/
Diferenciação Celular
/
Interferência de RNA
/
Linhagem Celular Tumoral
/
Proteínas Quinases Associadas a Fase S
/
Proliferação de Células
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Stomatology
Ano de publicação:
2007
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS