Recombinant adenovirus-mediated gene transfer to genitourinary epithelium in vitro and in vivo.
Recombinant adenovirus-mediated gene transfer to genitourinary epithelium in vitro and in vivo.
复制标题
重组腺病毒介导的体外和体内基因转移至泌尿生殖上皮。
作者：
C. Bass;G. Cabrera;A. Elgavish;B. Robert;G. Siegal;Anderson Sc;Maneval Dc;D. Curiel
膀胱移行细胞癌（TCC）与显性和隐性癌基因的特征性病变有关。在这些情况下，肿瘤发生的分子基础的理解使得可能的应用基因治疗策略的TCC。在这方面，通过尿道直接进入泌尿生殖道（GU）上皮的能力提供了实施这些各种基因治疗方法的实用手段。因此，我们探索了载体策略来实现GU上皮的直接体内转导。首先，使用表达萤火虫荧光素酶报告基因的重组腺病毒载体rAd-CMV-Luc转导三种人（HT 1197、HT 1376、T24）和一种小鼠（MBT-2）TCC细胞系。在这些研究中，发现报告基因表达在所有四种细胞系中均显著高于背景。值得注意的是，TCC细胞系HT 1197和HT 1376显示出与宫颈癌细胞系HeLa相当的表达水平，宫颈癌细胞系HeLa是先前显示出对重组腺病毒介导的基因转导高度敏感的细胞系。使用rAd-CMV-Luc的T24和MBT-2的体外时间过程显示T24系在转导后1天和MBT-2系在转导后3天达到峰值表达，可检测的表达水平持续至少7天。作为下一步，使用rAd-CMV-Luc转导源自GU上皮的人和小鼠原代组织。在该测定中，在两种情况下均观察到荧光素酶表达水平显著高于背景。（250字处删节）
Transitional cell carcinoma (TCC) of the bladder is associated with characterized lesions in dominant and recessive oncogenes. The understanding of the molecular basis of tumorigenesis in these instances makes possible the application of gene therapy strategies for TCC. In this regard, the ability to directly access the epithelium of the genitourinary (GU) tract via the urethra provides a practical means to implement these various gene therapy approaches. We thus explored vector strategies to accomplish direct in vivo transduction of GU epithelium. Initially, three human (HT 1197, HT 1376, T24) and one mouse (MBT-2) TCC cell lines were transduced using a recombinant adenoviral vector expressing the firefly luciferase reporter gene, rAd-CMV-Luc. In these studies, reporter gene expression was found to be significantly elevated above background for all four cell lines. Of note, the TCC cell lines HT 1197 and HT 1376 showed expression levels comparable with the cervical carcinoma cell line HeLa, a cell line previously shown to be highly susceptible to recombinant adenovirus-mediated gene transduction. An in vitro time course for T24 and MBT-2 using rAd-CMV-Luc showed peak expression 1 day after transduction for the T24 line and 3 days after transduction for the MBT-2 line, with detectable levels of expression persisting for at least 7 days. As a next step, human and mouse primary tissue deriving from the GU epithelium were transduced using rAd-CMV-Luc. In this assay, luciferase expression levels significantly above background were observed in both instances.(ABSTRACT TRUNCATED AT 250 WORDS)