An Optimized Telomerase-Specific Lentivirus for Optical Imaging of Tumors
An Optimized Telomerase-Specific Lentivirus for Optical Imaging of Tumors
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DOI：
10.1158/0008-5472.can-09-3841
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发表时间：
2010-04-01
期刊：
影响因子：
11.2
通讯作者：
Fang, Dian-Chun
中科院分区：
文献类型：
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作者：
Yu, Song-Tao;Yang, Yin-Bing;Fang, Dian-Chun
医学成像技术的进步，如超声、计算机断层扫描、磁共振成像和正电子发射断层扫描，在检测肿瘤方面取得了很大进展。然而，这些成像技术无法区分恶性肿瘤和良性肿瘤。最近发展的小动物肿瘤的光学成像提供了一种有用的方法来区分恶性肿瘤及其周围的正常组织。人端粒酶逆转录酶（hTERT）在大多数体细胞中通常是失活的，而它通常在许多癌细胞中被重新激活。在这项研究中，我们构建了一个慢病毒载体，表达绿色荧光蛋白（GFP）驱动的优化的hTERT启动子，以创建一个非侵入性的肿瘤特异性成像方法。发现这种优化的hTERT启动子的活性与SV 40和巨细胞病毒启动子的活性相等。体外实验表明，GFP仅在感染该慢病毒的端粒酶阳性肿瘤细胞中表达，而在端粒酶阴性肿瘤细胞或正常体细胞中没有GFP表达。我们还发现，通过使用电荷耦合器件（CCD）照相机，在瘤内注射这种慢病毒后24小时，可以观察到皮下端粒酶阳性肿瘤。相比之下，端粒酶阴性的肿瘤在瘤内注射后甚至30天也不能成像。这些结果表明，感染慢病毒含有这种优化的hTERT启动子可能是一个有用的诊断工具，用于实时可视化的宏观不可见的肿瘤组织，使用高灵敏度的CCD成像系统。Cancer Res; 70（7）; 2585-94. (C)2010年AACR。
Advances in medical imaging techniques, such as ultrasound, computed tomography, magnetic resonance imaging, and positron emission tomography, have made great progress in detecting tumors. However, these imaging techniques are unable to differentiate malignant tumors from benign ones. Recently developed optical imaging of tumors in small animals provides a useful method to distinguish malignant tumors from their surrounding normal tissues. Human telomerase reverse transcriptase (hTERT) is normally inactivated in most somatic cells, whereas it is commonly reactivated in many cancer cells. In this study, we constructed a lentiviral vector that expresses green fluorescent protein (GFP) driven by an optimized hTERT promoter to create a noninvasive tumor-specific imaging methodology. The activity of this optimized hTERT promoter was found to be equal to the activity of SV40 and cytomegalovirus promoters. In vitro experiments showed that GFP was only expressed in telomerase-positive tumor cells infected with this lentivirus, whereas there was no GFP expression in telomerase-negative tumor cells or normal somatic cells. We also found that subcutaneous telomerase-positive tumors could be visualized 24 hours after an intratumoral injection with this lentivirus by using a charge-coupled device (CCD) camera. In contrast, telomerase-negative tumors could not be imaged after an intratumoral injection even for 30 days. These results suggest that infection with lentivirus containing this optimized hTERT promoter might be a useful diagnostic tool for the real-time visualization of macroscopically invisible tumor tissues using a highly sensitive CCD imaging system. Cancer Res; 70(7); 2585-94. (C) 2010 AACR.