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Journal of Southern Medical University ; (12): 341-344, 2012.
Artigo em Chinês | WPRIM | ID: wpr-267604

RESUMO

<p><b>OBJECTIVE</b>To construct recombinant lentiviral vectors carrying Rheb gene and its mutant Rheb'D60K gene, and examine their expression in human liver cancer cells.</p><p><b>METHODS</b>Rheb gene was amplified by PCR to construct the recombinant plasmid LV31-Rheb-WT and LV31-Rheb-D60K. HEK-293 FT cells were contransfected with the recombinant lentiviral vector together with a lentiviral package plasmid to produce the lentiviral particles. The expression of PS6 protein was detected in the lentivirus-infected MCF-7 cells. The apoptosis of SK-HEP-1 cells transfected with LV31-Rheb-WT or LV31-Rheb-D60K was observed.</p><p><b>RESULTS</b>The recombinant LV31-Rheb-WT and LV31-Rheb-D60K vectors were confirmed by PCR and DNA sequencing. Western blotting showed that PS6 protein expression was increased in LV31-Rheb-WT-transfected cells while decreased in LV31-Rheb-D60K-transfected cells. LV31-Rheb-D60K-transfected SK-HEP-1 cells showed more obvious apoptosis after starvation than LV31-Rheb-WT-transfected cells.</p><p><b>CONCLUSION</b>Lentiviral vectors carrying Rheb gene and its mutant has been successfully constructed, which can be useful in further investigation of the role of Rheb gene in cancer cells.</p>


Assuntos
Humanos , Apoptose , Genética , Carcinoma Hepatocelular , Metabolismo , Patologia , Vetores Genéticos , Genética , Células HEK293 , Lentivirus , Genética , Metabolismo , Neoplasias Hepáticas , Metabolismo , Patologia , Células MCF-7 , Proteínas Monoméricas de Ligação ao GTP , Genética , Proteínas Mutantes , Genética , Neuropeptídeos , Genética , Proteína Enriquecida em Homólogo de Ras do Encéfalo , Proteínas Recombinantes , Genética , Transfecção
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