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A novel plasmid and SonoVue formulation plus ultrasound sonication for effective gene delivery in nude mice

机译：新型质粒和SonoVue配方加上超声超声处理可在裸鼠中有效传递基因

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Aims This study aimed to identify the effectiveness of gene therapy mediated by ultrasound-targeted SonoVue using the herpes simplex virus-thymidine kinase (TSV-TK) driven by the kinase insert domain receptor (KDR) promoter (KDR-TK) for the treatment of ovarian carcinomas in nude mice. The optimized conditions for gene transfection were also explored. Main methods In this study, we developed a novel technique to deliver a plasmid vector-carried gene into tumor xenografts in sixty nude mice. We first mixed plasmid DNA with SonoVue to form microbubbles and then treated the mice with ultrasound sonication to enhance plasmid gene transfection and expression in tumor xenografts. Key findings The data showed that injection of pBluescript-KDR-TK cDNA mixed with SonoVue into nude mice plus ultrasound sonication significantly (Group E) increased the transfection efficiency and expression of KDR-TK mRNA in tumor xenografts. The growth of tumor xenografts in nude mice was significantly suppressed in Group E compared to the other four control groups (Groups A, B, C, and D, namely, treatment with phosphate-buffered saline (PBS), KDR-TK + PBS, KDR-TK + SonoVue, KDR-TK + PBS + ultrasound sonication, respectively). TUNEL staining showed that SonoVue plus ultrasound sonication significantly induced apoptosis and reduced microvessel density (MVD). Significance This study revealed that the formulation of plasmid with SonoVue plus ultrasound could provide efficient gene delivery into tumor xenografts. Increased gene expression was observed in vivo, which effectively reduced the tumor growth and MVD of tumor xenografts and induced apoptosis in tumor cells. Future clinical trials are necessary to further analyze the relevance of this technique.
机译：目的本研究旨在确定由超声靶向的SonoVue介导的基因疗法的有效性，该疗法使用由激酶插入域受体（KDR）启动子（KDR-TK）驱动的单纯疱疹病毒胸苷激酶（TSV-TK）裸鼠卵巢癌。还探讨了基因转染的优化条件。主要方法在这项研究中，我们开发了一种新技术，可将携带质粒载体的基因传递到60只裸鼠的异种移植物中。我们首先将质粒DNA与SonoVue混合以形成微泡，然后用超声处理小鼠以增强质粒基因转染和在肿瘤异种移植物中的表达。关键发现数据显示，将与SonoVue混合的pBluescript-KDR-TK cDNA注入裸鼠并进行超声超声处理（E组）显着提高了肿瘤异种移植物中的转染效率和KDR-TK mRNA的表达。与其他四个对照组（A，B，C和D组，即用磷酸盐缓冲盐水（PBS），KDR-TK + PBS处理，E组）相比，E组裸鼠体内肿瘤异种移植物的生长受到显着抑制。 KDR-TK + SonoVue，KDR-TK + PBS +超声处理）。 TUNEL染色显示，SonoVue和超声超声处理可显着诱导细胞凋亡并降低微血管密度（MVD）。意义这项研究表明，用SonoVue加超声处理的质粒可以为肿瘤异种移植提供有效的基因传递。在体内观察到基因表达增加，这有效地降低了肿瘤异种移植物的肿瘤生长和MVD，并诱导了肿瘤细胞的凋亡。有必要进行进一步的临床试验，以进一步分析该技术的相关性。

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