Seo Ho Kyung, Seo Jeong Bin, Nam Jae-Kook, Jeong Kyung-Chae, Shin Seung-Pil, Kim In-Hoo, Lee Sang Don, Lee Sang-Jin
Center for Prostate Cancer, Gyeonggi-do, Biomedical Research Institute, Pusan National University and Research Institute for Convergence of Biomedical Science and Technology, Pusan.
Pusan National University and Biomedical Research Institute, Pusan National University and Research Institute for Convergence of Biomedical Science and Technology, Pusan.
Oncotarget. 2014 Jul 30;5(14):5615-23. doi: 10.18632/oncotarget.2151.
Survivin is a member of the inhibitors of apoptosis protein family. Here, we examined survivin expression and confirmed abundant survivin expression in bladder cancer cells. This expression pattern indicated that the transcriptional regulatory elements that control survivin expression could be utilized to discriminate cancer from normal cells. We therefore generated a novel adenovirus termed Ad5/35E1apsurvivinE4 with the following characteristics: 1) E1A and E4 protein expression was dependent on survivin promoter activity; 2) the green fluorescence protein gene was inserted into the genome under the control of the CMV promoter; 3) most of the E3 sequences were deleted, but the construct was still capable of expressing the adenovirus death protein with potent cytotoxic effects; and 4) the fiber knob was from serotype 35 adenovirus. As expected from the abundant survivin expression observed in bladder cancer cells, Ad5/35E1apsurvivinE4 replicated better in cancer cells than in normal cells by a factor of 106 to 102. Likewise, Ad5/35E1apsurvivinE4 exerted greater cytotoxic effects on all bladder cancer cell lines tested. Importantly, Ad5/35E1apsurvivinE4 inhibited the growth of Ku7-Luc orthotopic xenografts in nude mice. Taken together, Ad5/35E1apsurvivinE4 indicates that the survivin promoter may be utilized for the development of a replication-competent adenovirus to target bladder cancers.
生存素是凋亡抑制蛋白家族的成员之一。在此，我们检测了生存素的表达，并证实其在膀胱癌细胞中大量表达。这种表达模式表明，控制生存素表达的转录调控元件可用于区分癌细胞与正常细胞。因此，我们构建了一种新型腺病毒，命名为Ad5/35E1apsurvivinE4，其具有以下特点：1）E1A和E4蛋白的表达依赖于生存素启动子活性；2）绿色荧光蛋白基因在巨细胞病毒启动子的控制下插入基因组；3）大部分E3序列被删除，但该构建体仍能表达具有强大细胞毒性作用的腺病毒死亡蛋白；4）纤维结来自35型腺病毒。正如在膀胱癌细胞中观察到的大量生存素表达所预期的那样，Ad5/35E1apsurvivinE4在癌细胞中的复制能力比在正常细胞中强106至102倍。同样，Ad5/35E1apsurvivinE4对所有测试的膀胱癌细胞系都具有更强的细胞毒性作用。重要的是，Ad5/35E1apsurvivinE4抑制了裸鼠体内Ku7-Luc原位异种移植物的生长。综上所述，Ad5/35E1apsurvivinE4表明生存素启动子可用于开发一种有复制能力的腺病毒来靶向膀胱癌。
