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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>
Assuntos
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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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