Construction of Recombinant Newcastle Disease Virus for Oncolytic Therapy of Cancer
会议名称：《“细胞活动 生命活力”——中国细胞生物学学会全体会员代表大会暨第十二次学术大会》
会议日期：2011年
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
关 键 词：Newcastle disease virus Reverse genetic technology Oncolytic virus Cancer
摘 要：Newcastle disease virus(NDV),a naturally oncolytic virus,is known as a potential agent for cancer ***, we constructed a reverse genetic system based on Italien strain which is an oncolytic and virulence strain of NDV. Using the reverse genetic system,two recombinant NDV carrying EGFP and firefly luciferase genes were *** the proliferation kinetic and the anti-tumor activity of recombinant viruses were evaluated in vitro and in ***:To generate the recombinant NDV,a minigenome plasmid,three helper plasmids and genome plasmid were *** complete coding sequence of EGFP or firefly luciferase was inserted into the non-coding region between HN and F *** helper plasmids and genome plasmid were co-transfected into BSR-T7/5 cells to rescue the recombinant NDV To evaluate the propagation and anti-tumor characteristics,several cell lines were infected with recombinant NDV and NDV Italien,and the viral titer and cell viability were measured in different time *** evaluate the anti-tumor activity in vivo,tumor-bearing nude mice were infected with the recombinant virus through intratumoral or intravenous injection,the exogenous gene expression were observed by living image system and the tumor volume and survival time were ***:Recombinant NDV were collected and purified at 5 day post transfection and identified by genome *** data showed the insertion of exogenous reporter gene did not affect the viral propagation efficiency,and the recombinant NDV kept the characteristics of tumor-selective replication and the sensitivity to type I interferon similar as wild-type NDV Living image showed the recombinant NDV replicated in tumor tissue of nude mice ***,the recombinant NDV prolonged the survival time of nude mice significantly by intratumoral injection(P0.05). Conclusion:We generated the recombinant NDV with reporter gene based on NDV Italien strain for the first time which kept the propagation pr