[Silencing of COX-2 in nasopharyngeal carcinoma cells with a shRNAmir lentivirus vector].
OBJECTIVE
To construct a miR-155-based lentivirus vector to induce cyclooxygenase-2 gene silencing in nasopharyngeal carcinoma (NPC) cells by expressing anti-COX-2 shRNAmir.


METHODS
miR-155-based anti-COX-2 shRNAmir template was synthesized and inserted into pLVTHM plasmid. The recombinant pLVTHM/shRNAmir was transfected into 293FT cells for packaging the lentivirus vector. After infection with the lentivirus vector, the GFP-positive cells were screened by flow cytometry, and COX-2 mRNA level was detected by RT-PCR.


RESULTS
Restriction digestion and DNA sequencing confirmed successful construction of the anti-COX-2 vector pLVTHM/shRNAmir. A subline of C666-1 cells was established after infection with the lentivirus vector, and the COX-2 expression in the cells was stably silenced.


CONCLUSIONS
The shRNAmir lentivirus vector constructed may serve as an effective COX-2 inhibitor, which may facilitate future studies of gene therapy of NPC.
Li
Gang
G
Department of Otolaryngology, Nanfang Hospital, Southern Medical University, Guangzhou, China. climber1999y@yahoo.com.cn
Li
Xiang-Ping
XP
Jiang
Li
L
Lu
Juan
J
Liu
Xiong
X
Chen
Shun-Jin
SJ
D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics; D009303:Nasopharyngeal Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
C524942:MIRN155 microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human