Department of Neurosurgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China; Institute of Ultrasound Imaging, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.
Department of Neurosurgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China.
Cancer Lett. 2018 Apr 1;418:147-158. doi: 10.1016/j.canlet.2018.01.035. Epub 2018 Jan 12.
Our previous studies have demonstrated that focused ultrasound (FUS) combined with DNA-loaded microbubbles (MBs) can induce noninvasive, reversible, local disruption of the blood brain barrier (BBB) and enable targeted exogenous gene transfer into the central nervous system. However, due to low gene loading or the absence of positive targeting, to date, there has been no therapeutic effect of MBs combined with FUS in tumor treatment. In the current study, we adopted a phospholipid complex that exhibited sufficient gene loading and peptide-mediated targeting to delay glioma growth. First, we bound MBs to shBirc5-lipo-NGR, which performed the dual function of tumor cell targeting and effective gene loading. Next, we demonstrated that FUS-aided MB-shBirc5-lipo-NGR exhibited a higher transfection efficiency compared with the control group. Finally, we evaluated the silencing effect of shBirc5 using an apoptosis assay, real time-polymerase chain reaction (PCR), western blotting (WB) in vitro and a volume measurement survival analysis in vivo. The experimental group exhibited a significant therapeutic effect, while the FUS-only, MB-shBirc5-lipo-NGR-only and FUS-aided MB-shControl-lipo-NGR groups displayed no changes in tumor growth or survival time (P < .01). Consequently, our study indicated that MB-shBirc5-lipo-NGR combined with FUS is a promising new RNA interference technique for the treatment of glioma.
我们之前的研究表明，聚焦超声（FUS）联合载 DNA 的微泡（MBs）可以诱导非侵入性、可逆的血脑屏障（BBB）局部破坏，并使靶向的外源基因转移到中枢神经系统。然而，由于载基因量低或缺乏阳性靶向，迄今为止，MBs 联合 FUS 在肿瘤治疗中尚未产生治疗效果。在本研究中，我们采用了一种具有足够载基因量和肽介导靶向性的磷脂复合物来延迟神经胶质瘤的生长。首先，我们将 MBs 与 shBirc5-lipo-NGR 结合，使其具有肿瘤细胞靶向和有效载基因的双重功能。接下来，我们证明 FUS 辅助的 MB-shBirc5-lipo-NGR 与对照组相比具有更高的转染效率。最后，我们通过体外凋亡检测、实时聚合酶链反应（PCR）、western blot（WB）以及体内体积测量生存分析来评估 shBirc5 的沉默效果。实验组表现出显著的治疗效果，而仅 FUS、仅 MB-shBirc5-lipo-NGR 和 FUS 辅助的 MB-shControl-lipo-NGR 组的肿瘤生长或存活时间没有变化（P<0.01）。因此，我们的研究表明，MB-shBirc5-lipo-NGR 联合 FUS 是一种有前途的治疗神经胶质瘤的新 RNA 干扰技术。
