[Effective inhibition of EB virus-encoded latent membrane protein-1 by siRNA in EB virus (+) nasopharyngeal carcinoma cell]

Gang Li, Xiang-ping Li, Ying Peng, Xiong Liu, Xiao-hua Li
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke za Zhi, Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2005, 40 (6): 406-10

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.

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