组织切片模型系统在测定溶瘤腺病毒感染乳腺癌中的效用。
The utility of a tissue slice model system to determine breast cancer infectivity by oncolytic adenoviruses.
机构信息
Department of Surgery, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130, USA.
出版信息
J Surg Res. 2010 Oct;163(2):270-5. doi: 10.1016/j.jss.2010.03.072. Epub 2010 Apr 24.
BACKGROUND
Due to advances in viral design, oncolytic adenoviruses have emerged as a promising approach for treatment of breast cancer. Tumor tissue slices offer a stringent model system for preclinical evaluation of adenovirus therapies, since the slices retain a morphology and phenotype that more closely resembles the in vivo setting than cell line cultures, and this system has been shown to have utility in the evaluation of viral infectivity and replication. In this study, we evaluated the efficacy of viral infection and replication using a tropism-modified oncolytic adenovirus.
METHODS
Breast tumor tissue slices were infected with a tropism-modified oncolytic adenovirus, and a wild-type adenovirus for comparison. Efficiency of infection was evaluated using fluorescent microscopy, as the viruses used have been modified to express red fluorescent protein. Replication of the viruses was evaluated with quantitative real-time polymerase chain reaction (PCR) to assay viral E4 genome copy number, a surrogate indicator for the number of virions. The breast tumor tissue slices were evaluated for the expression of CD46 expression by immunohistochemistry.
RESULTS
Infection and replication of our tropism modified oncolytic virus has been observed in the breast cancer tissue slice model system and is comparative to wild-type virus. A qualitative increase in the number of cells showing red fluorescent protein (RFP) expression was observed correlating with increasing multiplicity of infection. Higher relative infectivity of the virus was observed in tumor tissue compared with normal breast tissue. Replication of the virus was demonstrated through increases in E4 copy number at 48 and 72 h after infection in human breast tumor slices.
CONCLUSIONS
We have shown that a tropism modified oncolytic adenovirus can infect and replicate in breast cancer tissue slices, which may be an important preclinical indicator for its therapeutic utility.
背景
由于病毒设计的进步，溶瘤腺病毒已成为治疗乳腺癌的一种很有前途的方法。肿瘤组织切片为腺病毒治疗的临床前评估提供了一个严格的模型系统，因为这些切片保留了更接近体内环境的形态和表型，而不是细胞系培养，并且已经证明该系统在评估病毒感染力和复制方面具有实用性。在这项研究中，我们使用一种靶向修饰的溶瘤腺病毒评估了病毒感染和复制的效果。
方法
将靶向修饰的溶瘤腺病毒和野生型腺病毒用于感染乳腺肿瘤组织切片。使用荧光显微镜评估感染效率，因为所使用的病毒已被修饰以表达红色荧光蛋白。使用定量实时聚合酶链反应 (PCR) 评估病毒 E4 基因组拷贝数来评估病毒的复制，这是病毒体数量的替代指标。通过免疫组织化学评估乳腺肿瘤组织切片中 CD46 表达的情况。
结果
在乳腺癌组织切片模型系统中观察到我们的靶向修饰溶瘤病毒的感染和复制，与野生型病毒相当。随着感染复数的增加，观察到表达红色荧光蛋白 (RFP) 的细胞数量增加，表明存在定性增加。与正常乳腺组织相比，病毒在肿瘤组织中的相对感染力更高。在感染后 48 和 72 小时，通过 E4 拷贝数的增加证明了病毒的复制。
结论
我们已经表明，一种靶向修饰的溶瘤腺病毒可以感染和在乳腺癌组织切片中复制，这可能是其治疗用途的重要临床前指标。
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