Liang Qianwa, Dmitriev Igor, Kashentseva Elena, Curiel David T, Herschman Harvey R
Department of Biological Chemistry, Molecular Biology Institute, David Geffen School of Medicine, Los Angeles, CA USA.
Mol Imaging Biol. 2004 Nov-Dec;6(6):385-94. doi: 10.1016/j.mibio.2004.09.001.
To demonstrate that non-invasive bioluminescent imaging can monitor restricted expression from a nonreplicating adenovirus in which the cyclooxygenase-2 (COX-2) promoter drives firefly luciferase.
Adenovirus in which the COX-2 promoter drives the firefly luciferase imaging gene was injected intratumorally into xenografts that express relatively low and relatively high levels of COX-2. Adenovirus that expresses Renilla Luciferase from the cytomegalovirus early promoter was co-injected, to normalize for injection, leakage, vascularization, etc. COX-2 restricted firefly luciferase and global Renilla Luciferase activities were measured by optical imaging techniques both in vivo and in isolated tissues.
Dramatic reduction in hepatic luciferase expression after intravenous viral injection can be imaged non-invasively in living animals. Following intratumoral injection, luciferase levels in tumor xenografts that express differing endogenous COX-2 levels reflect the luciferase levels observed when these cells are infected in cell culture. Essentially no luciferase expression is observed in liver following intratumoral injection.
Both tissue restricted expression and transcriptional redirection to tumors expressing COX-2 can be imaged non-invasively following injection of Adenovirus expressing firefly luciferase from the COX-2 promoter.
证明非侵入性生物发光成像可监测由环氧合酶-2（COX-2）启动子驱动萤火虫荧光素酶的非复制性腺病毒的限制性表达。
将COX-2启动子驱动萤火虫荧光素酶成像基因的腺病毒瘤内注射到表达相对低水平和相对高水平COX-2的异种移植瘤中。同时共注射从巨细胞病毒早期启动子表达海肾荧光素酶的腺病毒，以校正注射、渗漏、血管形成等因素。通过光学成像技术在体内和离体组织中测量COX-2限制性萤火虫荧光素酶和整体海肾荧光素酶活性。
静脉注射病毒后肝脏中荧光素酶表达的显著降低可在活体动物中进行非侵入性成像。瘤内注射后，表达不同内源性COX-2水平的肿瘤异种移植瘤中的荧光素酶水平反映了这些细胞在细胞培养中感染时观察到的荧光素酶水平。瘤内注射后肝脏中基本未观察到荧光素酶表达。
注射由COX-2启动子表达萤火虫荧光素酶的腺病毒后，组织限制性表达和向表达COX-2的肿瘤的转录重定向均可进行非侵入性成像。
