Oncolysis and suppression of tumor growth by a GFP-expressing oncolytic adenovirus controlled by an hTERT and CMV hybrid promoter
Oncolysis and suppression of tumor growth by a GFP-expressing oncolytic adenovirus controlled by an hTERT and CMV hybrid promoter
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DOI：
10.1038/sj.cgt.7700944
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发表时间：
2006-07-01
影响因子：
6.4
通讯作者：
Fang, B.
中科院分区：
文献类型：
--
作者：
Davis, J. J.;Wang, L.;Fang, B.
溶瘤病毒疗法的挑战之一是不能在治疗期间容易地跟踪或监测病毒活性。在这里，我们描述的Ad/hTC-GFP-E1，溶瘤病毒的转基因GFP和E1 A都在一个合成的启动子（hTC）的控制下的构建和功能特性。该启动子由来自人端粒酶逆转录酶启动子和最小巨细胞病毒（CMV）早期启动子的序列组成。通过Western blot和荧光显微镜分析证实E1 A和GFP的肿瘤特异性表达，并通过结晶紫染色和细胞活力测定证实肿瘤特异性细胞毒性。在敏感细胞系中，病毒复制和肿瘤细胞溶解发生在感染复数（MOI）低至100个病毒颗粒/细胞。在正常人成纤维细胞中未观察到明显的细胞毒性作用，即使MOI超过2000 vp。溶瘤载体的存在很容易在体外和体内可视化和定量，与病毒复制相关。病灶内给药的病毒皮下H1299（NSCLC）肿瘤异种移植显着抑制肿瘤生长，并提供了生存的好处。总之，这些结果证明表达hTERT特异性转基因的hTERT特异性溶瘤腺病毒适用于癌症治疗。
One of the challenges of oncolytic virotherapy is the inability to easily track or monitor virus activity during treatment. Here we describe the construction and functional characterization of Ad/hTC-GFP-E1, an oncolytic virus whose transgenes GFP and E1A are both under the control of a synthetic promoter (hTC). This promoter consists of sequences from the human telomorase reverse transcriptase promoter and a minimal cytomegalovirus (CMV) early promoter. The tumor-specific expression of E1A and GFP was demonstrated by Western blot and fluorescent microscope analyses, and the tumor-specific cytotoxicity by crystal-violet staining and cell viability assays. Viral replication and tumor cell lysis occured at multiplicities of infection (MOI) as low as 100 viral particles per cell in sensitive cell lines. No overt cytotoxic effect was observed in normal human fibroblasts, even at MOIs over 2000 vp. The presence of oncolytic vector was easily visualized and quantitated in vitro and in vivo, in correlation with viral replication. Intralesional administration of the virus into subcutaneous H1299 (NSCLC) tumor xenografts significantly suppressed tumor growth and provided a survival benefit. Together, these results demonstrate that an hTERT-specific oncolytic adenovirus expressing an hTERT-specific transgene is applicable for cancer therapy.