Effect of inhibition of EBV-encoded latent membrane protein-1 by small interfering RNA on EBV-positive nasopharyngeal carcinoma cell growth

Gang Li, Xiang-ping Li, Ying Peng, Xiong Liu, Xiao-hua Li
Di 1 Jun Yi da Xue Xue Bao, Academic Journal of the First Medical College of PLA 2004, 24 (3): 241-6

OBJECTIVE: To evaluate the feasibility of using small interfering RNA (siRNA) for selective inhibiting latent membrane protein-1 (LMP1) expression in Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC) cell line C611, and observe the effects of LMP1 gene silencing on the NPC cell growth.

METHODS: Four synthesized double-strand siRNA were respectively transfected into NPC cells, using Oligofectamine reagent. The subsequent changes in LMP1 mRNA level were determined by semi-quantitative reverse transcriptase (RT)-PCR. Flow cytometry and MTT assay were employed to examine the alterations in cell cycle and cell proliferation.

RESULTS: The most effective sequence was identified among the 4 candidate siRNAs, and its single dose caused nearly 90% loss of LMP1 mRNA in C611 cells, with sustained specific inhibition for 96 h following a re-transfection. LMP1 siRNA treatment resulted in cell cycle arrest at G(0)-G(1) phase, accompanied by a reduction of cell proliferation by 33%, whereas EBV-negative NPC cells appeared unaffected.

CONCLUSIONS: EBV-encoded LMP-1 is vulnerable to RNA interference and selective inhibition of LMP1 suppresses the proliferation of EBV-positive NPC cells, a finding that sheds light on the possible use of RNA interference in further investigations of LMP1 and for therapeutic purposes of EBV-related NPC.

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