A novel gene expression system for detecting viable bladder cancer cells
A novel gene expression system for detecting viable bladder cancer cells
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用于检测存活膀胱癌细胞的新型基因表达系统
DOI：
10.3892/ijo.2012.1417
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发表时间：
2012-07-01
影响因子：
5.2
通讯作者：
Kumon, Hiromi
中科院分区：
文献类型：
--
作者：
Ueki, Hideo;Watanabe, Masami;Kumon, Hiromi
开发了一种新型转录系统，可以强有力地增强癌症特异性基因的表达。在该系统中，hTERT 启动子驱动的基因表达通过先进的两步转录扩增 (TSTA) 得到增强。该构建体用于开发检测膀胱癌细胞的新系统。目前的研究通过检查各种膀胱癌细胞系中的癌症特异性基因转录来评估先进的 TSTA 系统。与之前的一步或传统 TSTA 表达系统相比，该系统显着增强了膀胱癌细胞系中癌症特异性荧光素酶基因的表达。增强的倍数增益与细胞系的端粒酶活性显着相关。构建了绿色荧光蛋白 (GFP) 基因编码质粒载体，其中先进的 TSTA 增强了 hTERT 启动子驱动转录，以利用该系统对活膀胱癌细胞进行成像和检测。先进的 TSTA-hTERT-GFP 质粒成功诱导癌症特异性基因表达，在人膀胱癌细胞系中显示出强大的 GFP 表达，但在正常膀胱尿路上皮细胞中没有可见的 GFP 表达。具有CMV启动子的对照GFP质粒在正常膀胱细胞和癌细胞中均产生GFP表达。先进的 TSTA-hTERT-GFP 质粒允许在含有 10 倍和 100 倍正常膀胱尿路上皮细胞的混合细胞培养物中选择性地观察活的人膀胱癌细胞。这些发现表明先进的 TSTA-hTERT 表达系统是检测存活膀胱癌细胞的宝贵工具。目前的系统可用于体外检测尿液中的膀胱癌细胞和体内播散的其他类型的癌细胞。
A novel transcriptional system was developed that can robustly enhance cancer-specific gene expression. In the system, hTERT promoter-driven gene expression was enhanced by an advanced two-step transcriptional amplification (TSTA). This construct was used to develop a novel system for detection of bladder cancer cells. The current study evaluated the advanced TSTA system by examining the cancer-specific gene transcription in various bladder cancer cell lines. The system significantly enhanced cancer-specific luciferase gene expression in the bladder cancer cell lines in comparison to the previous expression system of one-step or conventional TSTA. The fold gain of the enhancement was significantly correlated to the telomerase activity of the cell lines. A green fluorescent protein (GFP) gene encoding plasmid vector was constructed where hTERT promoter-driving transcription is enhanced by the advanced TSTA to utilize the system for the imaging and detection of viable bladder cancer cells. The advanced TSTA-hTERT-GFP plasmid successfully induced cancer-specific gene expression, showing robust GFP expression in human bladder cancer cell lines, but no visible GFP expression in normal bladder urothelial cells. The control GFP plasm id with a CMV promoter yielded GFP expression in both normal bladder cells and cancer cells. The advanced TSTA-hTERT-GFP plasmid allowed selective visualization of viable human bladder cancer cells in mixed cell culture containing 10- and 100-fold more normal bladder urothelial cells. These findings indicate that the advanced TSTA-hTERT expressional system is a valuable tool for detecting viable bladder cancer cells. The current system can be applied for in vitro detection of bladder cancer cells in urine and other types of cancer cells disseminated in vivo.