[Effective inhibition of EB virus-encoded latent membrane protein-1 by siRNA in EB virus (+) nasopharyngeal carcinoma cell].
OBJECTIVE
To evaluate the application of RNA interference in the study of latent membrane protein-1 (LMP-1) in EB virus-positive nasopharyngeal carcinoma (NPC) cell line C611. Observe the effects of LMP-1 silencing on NPC cell growth.


METHODS
Four synthesized small interference RNA (siRNA) were transfected into NPC cell using Oligofectamine reagent. LMP-1 mRNA level was determined by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). Flow cytometry and MTT assay were employed to analyze the effects on cell cycle and proliferation.


RESULTS
The most effective sequence found out among the 4 candidates. Single dose of this siRNA caused nearly 90% loss of LMP-1 mRNA in C611 cell. The specific inhibition could last for 96 hours if re-transfection was preformed. LMP-1 siRNA treatment resulted in cell cycle arrest at G0-G1 stage in C611, accompany with a reduction of cell proliferation by 32.9%. While EBV-negative NPC cells appeared unaffected.


CONCLUSIONS
These results provided solid testimony that EBV-encoded latent membrane protein-1 was vulnerable to RNA interference and, selective inhibition of LMP-1 had anti-proliferation effect on NPC cell. RNA interference could be a powerful tool in further investigations of LMP-1 and a novel therapeutic strategy for EBV-related NPC patients.
Li
Gang
G
Deparrtment of Otorhinolaryngology Head and Neck Surgery, Nanfang Hospital, Nanfang Medical University, Guangzhou 510515, China.
Li
Xiang-ping
XP
Peng
Ying
Y
Liu
Xiong
X
Li
Xiao-hua
XH
D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004854:Herpesvirus 4, Human / Q000378:metabolism; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000235:genetics / Q000473:pathology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics*
C037662:EBV-associated membrane antigen, Epstein-Barr virus; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014763:Viral Matrix Proteins