Suppression Of Epstein-Barr Virus-encoded Latent Membrane Protein 1 By AAV Vector Mediate RNA Interference Inbibits Metastasis Potential Of Nasopharyngeal Cells In Vivo And In Vitro

Objective:1. To evaluate the feasibility of recombinant adeno-associated virus mediated RNA interference on LMP-1 and its inhibting efficiency on taget gene.2. To investigate the contribution of EBV-encoded LMP-1 in vivo on NPC cell growth and metastasis.3. To evaluate the significance of recombinant adeno-associated virus mediated shRNA-LMP-1 interference gene therapy on NPC.Methods:To construct recombinant adeno-associated virus (rAAV) vector which mediating specific small hairpin RNA (shRNA) on nasopharyngeal carcinoma Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP-1). The inhibition efficiency on LMP-1 was assessed by PT-PCR. The biological effect after "gene silencing" of LMP-1 on NPC cells(C666-l) of cell cycle, cell proliferation and metastasis ability in vitro were performed by MTT array, flow cytometry, cells mobility and the ability for matrigel transmembrane migration.EBV positive NPC ceIls(C666-l) which treated by RNAi on LMP-1 in vitro was inoculated by direct injection into the liver via laparotomy under direct visualization to establish the animal model of NPC xenografted into liver and liver to lung metastasis. The biological effect and its mechanism after "gene silencing" of LMP-1...