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
in Chinese
| WPRIM
| ID: wpr-268830
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>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
RNA, Messenger
/
Transfection
/
Nasopharyngeal Neoplasms
/
Lentivirus
/
Eukaryotic Initiation Factor-4G
/
RNA, Small Interfering
/
RNA Interference
/
Cell Line, Tumor
/
Genetic Vectors
/
Genetics
Limits:
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2009
Type:
Article
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