Department of Biology, University of Rochester, Rochester, New York 14627, USA.
Mol Ther. 2012 Feb;20(2):347-55. doi: 10.1038/mt.2011.215. Epub 2011 Oct 18.
Rad51 protein is overexpressed in a wide range of human cancers. Our previous in vitro studies demonstrated that a construct comprised Rad51 promoter driving expression of the diphtheria toxin A gene (pRad51-diphtheria toxin A (DTA)) destroys a variety of human cancer cell lines, with minimal to no toxicity to normal human cells. Here we delivered Rad51 promoter-based constructs in vivo using linear polyethylenimine nanoparticles, in vivo jetPEI, to visualize and treat tumors in mice with HeLa xenografts. For tumor detection, we used pRad51-Luc, a construct containing the firefly luciferase under the Rad51 promoter, administered by intraperitoneal (IP) injection. Tumors were detected with an in vivo bioluminescent camera. All mice with cancer displayed strong bioluminescence, while mice without cancer displayed no detectable bioluminescence. Treatment with pRad51-DTA/jetPEI decreased tumor mass of subcutaneous (SC) and IP tumors by sixfold and fourfold, respectively, along with the strong reduction of malignant ascites. Fifty percent of the mice with SC tumors were cancer-free after six pRad51-DTA/jetPEI injections, and for the mice with IP tumors, mean survival time increased by 90% compared to control mice. This study demonstrates the clinical potential of pRad51-based constructs delivered by nanoparticles for the diagnostics and treatment of a wide range of cancers.
Rad51 蛋白在多种人类癌症中过度表达。我们之前的体外研究表明，由 Rad51 启动子驱动白喉毒素 A 基因表达的构建体（pRad51-白喉毒素 A（DTA））可破坏多种人类癌细胞系，对正常人类细胞几乎没有毒性。在这里，我们使用线性聚乙烯亚胺纳米粒子（in vivo jetPEI）在体内传递基于 Rad51 启动子的构建体，以可视化和治疗带有 HeLa 异种移植物的小鼠肿瘤。为了进行肿瘤检测，我们使用了 pRad51-Luc，这是一种含有 Rad51 启动子下的萤火虫荧光素酶的构建体，通过腹腔内（IP）注射给药。使用体内生物发光相机检测肿瘤。所有患有癌症的小鼠均显示出强烈的生物发光，而没有癌症的小鼠则未检测到生物发光。用 pRad51-DTA/jetPEI 治疗可使皮下（SC）和 IP 肿瘤的肿瘤质量分别减少六倍和四倍，同时恶性腹水也明显减少。在接受六次 pRad51-DTA/jetPEI 注射后，有 50%的 SC 肿瘤小鼠无癌症，而 IP 肿瘤小鼠的中位生存时间比对照组小鼠延长了 90%。这项研究表明，纳米粒子传递的基于 pRad51 的构建体在诊断和治疗广泛的癌症方面具有临床潜力。
