Establishment of a stable nasopharyngeal carcinoma cell line with lentivirus-mediated RNA interference for EIF4G1 gene silencing / 南方医科大学学报
Journal of Southern Medical University
; (12): 844-851, 2009.
Article
de Zh
| WPRIM
| ID: wpr-268830
Bibliothèque responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To establish a nasopharyngeal carcinoma (NPC) cell line with stable EIF4G1 gene silencing induced by small interfering RNA (siRNA).</p><p><b>METHODS</b>The EIF4G1 mRNA levels in 8 NPC cell lines including 5-8F, 6-10B, C666-1, CNE1, CNE2, HNE1, HONE1, and SUNE1 were detected by fluorescence quantitative RT-PCR (QRT-PCR). The recombinant lentivirus shRNA expression plasmid targeting EIF4G1 gene was packaged into mature lentivirus by 293FT cells and used to infect 5-8F cells. After blasticidin selection of NPC cells with constant expression of the EIF4G1-siRNA, the efficiency of EIF4G1 mRNA expression interference was determined using QRT-PCR.</p><p><b>RESULTS</b>The 8 NPC cell lines showed differential expression of EIF4G1 mRNA, among which 5-8F cells had the highest EIF4G1 expression. The recombinant lentivirus plasmid pLenti6/BLOCK-iT-DEST/EIF4G1-shRNA was successfully constructed and verified by PCR and sequencing. The EIF4G1 mRNA level of 5-8F cells infected with shRNA-EIF4G1 lentivirus was significantly reduced as compared with the negative control and the blank control cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus vector pLenti6/BLOCK- iT-DEST/EIF4G1-shRNA we constructed results in marked downregulation of EIF4G1 mRNA expression and constant expression of EIF4G1-siRNA after infection of 5-8F cells.</p>
Texte intégral:
1
Base de données:
WPRIM
Sujet principal:
ARN messager
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Transfection
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Tumeurs du rhinopharynx
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Lentivirus
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Facteur-4G d'initiation eucaryote
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Petit ARN interférent
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Interférence par ARN
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Lignée cellulaire tumorale
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Vecteurs génétiques
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Génétique
Limites:
Humans
Langue:
Zh
Journal:
Journal of Southern Medical University
Année:
2009
Type de document:
Article