Experimental study on protective effect of small interfering RNA-induced Atrogin-1 gene silencing on muscle cell malnutrition / 中华外科杂志
Chinese Journal of Surgery
;
(12): 705-708, 2009.
Artigo
em Chinês
| WPRIM
| ID: wpr-280596
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the protective effect of Atrogin-1 gene silencing via RNA interference technique on a model of muscle cell malnutrition.</p><p><b>METHODS</b>Sequences of five target Atrogin-1 siRNA and the control were selected and synthesized and cloned to vector pBS-hU6-I and then to vector FG12. The length and rightness of the sequences were confirmed. The recombinant FG12 vectors were cotransfected along with pRSVREV, pMDLg/pRRE and pHCMV-G into 293T cells to package lentivirus particles, with which C2C12 cells were infected. The infected C2C12 cells were cultured and differentiated to form myotubes before TNF-alpha was added to induce malnutrition. Expressed products of Atrogin-1 of myotubes were identified by real time PCR and Western blot methods. Myotubes were observed and photographed directly in culture plate without fixation.</p><p><b>RESULTS</b>The length and sequences of inserted DNA were right. Compared with the RNA interferencing group, significant atrophy and upregulated expression of Atrogin-1 of myotubes treated by TNF-alpha was found in the control group.</p><p><b>CONCLUSION</b>Atrogin-1 gene silencing could be used to inhibit malnutrition of muscle cells caused by TNF-alpha. Atrogin-1 could be an ideal target in the treatment of cancer cachexia.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Farmacologia
/
Transfecção
/
Diferenciação Celular
/
Células Cultivadas
/
Fator de Necrose Tumoral alfa
/
Lentivirus
/
Inativação Gênica
/
Biologia Celular
/
Células Musculares
/
RNA Interferente Pequeno
Tipo de estudo:
Estudo prognóstico
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Surgery
Ano de publicação:
2009
Tipo de documento:
Artigo
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