Xu Jinfeng, Zeng Xinxin, Liu Yingying, Luo Hui, Wei Zhanghong, Liu Huiyu, Zhou Yuli, Zheng Hairong, Zhou Jie, Tan Guanghong, Yan Fei
Department of Ultrasonography, Shenzhen People's Hospital, Second Clinical Medical College of Jinan University, Shenzhen, 518020, China.
Paul C. Lauterbur Research Center for Biomedical Imaging, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
Tumour Biol. 2016 Jul;37(7):8609-19. doi: 10.1007/s13277-015-4681-7. Epub 2016 Jan 6.
Ultrasound-targeted microbubble destruction (UTMD) has become a novel gene/drug delivery method in cancer therapeutic application. However, the gene transfection efficiency mediated by UTMD is still unsatisfactory. Here, we introduced iRGD/CCR2 dual-targeted cationic microbubbles (MBiRGD/CCR2) which was modified with PEI-600 and coated with iRGD peptides and anti-CCR-2 antibodies. It showed that MBiRGD/CCR2 had a 25.83 ± 1.57 mV surface zeta potential and good stability. The experiments in vitro showed MBiRGD/CCR2 had higher binding efficiency with both bEnd.3 cells and MCF-7 cells than that of iRGD or CCR2 single-targeted cationic microbubbles (MBiRGD or MBCCR2) (P < 0.05 for both). Agarose gel electrophoresis assay showed that MBiRGD/CCR2 can effectively load pGPU6/GFP/Neo-shAKT2 plasmid DNA. Compared with the plain MBs (MBcontrol) or single-targeted cationic MBs including MBiRGD and MBCCR2 (P < 0.05 for all), the dual-targeted cationic MBiRGD/CCR2 groups had higher gene transfection efficiency under US exposure. It showed that the dual-targeted cationic MBiRGD/CCR2 has a potential value to be used as an ultrasound imaging probe for ultrasound image-guided tumor gene therapy.
超声靶向微泡破坏（UTMD）已成为癌症治疗应用中的一种新型基因/药物递送方法。然而，UTMD介导的基因转染效率仍不尽人意。在此，我们引入了用PEI-600修饰、包被iRGD肽和抗CCR-2抗体的iRGD/CCR2双靶向阳离子微泡（MBiRGD/CCR2）。结果显示，MBiRGD/CCR2具有25.83±1.57 mV的表面zeta电位和良好的稳定性。体外实验表明，MBiRGD/CCR2与bEnd.3细胞和MCF-7细胞的结合效率均高于iRGD或CCR2单靶向阳离子微泡（MBiRGD或MBCCR2）（两者P均<0.05）。琼脂糖凝胶电泳分析表明，MBiRGD/CCR2能够有效负载pGPU6/GFP/Neo-shAKT2质粒DNA。与普通微泡（MBcontrol）或包括MBiRGD和MBCCR2在内的单靶向阳离子微泡相比（所有P均<0.05），双靶向阳离子MBiRGD/CCR2组在超声照射下具有更高的基因转染效率。结果表明，双靶向阳离子MBiRGD/CCR2作为超声图像引导肿瘤基因治疗的超声成像探针具有潜在应用价值。
