Ultrasound-mediated microbubble destruction enhances gene transfection in pancreatic cancer cells
Ultrasound-mediated microbubble destruction enhances gene transfection in pancreatic cancer cells
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DOI：
10.1007/s12325-008-0051-9
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发表时间：
2008-05-01
影响因子：
3.8
通讯作者：
Wu, Chang-Jun
中科院分区：
文献类型：
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作者：
Wang, Jun-Feng;Wang, Jia-Bei;Wu, Chang-Jun
前言：本研究的目的是确定超声暴露联合微泡破坏是否可以增强人胰腺癌细胞（PANC-1）的非病毒基因传递。方法：分为4个实验组：P组，质粒单独；P+M组，质粒和微泡；P+U组，质粒和超声；P+U+M组，质粒超声加微泡。编码增强型绿色荧光蛋白（pEGFP）的质粒DNA与市售超声微泡造影剂(SonoVue (R)；博莱科诊断公司，米兰，意大利)在集团P+M和集团P+U+M。在不同条件下将DNA和DNA +微泡的不同组合加入培养的PANC-1细胞中。转染效率和细胞活力通过FACS分析（Becton Dickinson, San Jose， CA， USA）、共聚焦激光扫描显微镜和台盼蓝染色进行评估。结果：超声暴露微泡与其他组（P+U+M组，21.4%+/- 3.16%；P组，2.9%+/- 0.45%；P+M组，3.1%+/- 0.51%；P+U组，6.1%+/- 1.27%;P < 0.01）相比，能显著提高报告基因的表达。P+U+M组PANC-1细胞活力与其他组比较差异无统计学意义（P < 0.05）。结论：我们的体外研究结果表明，超声介导的微泡破坏有可能促进有效的基因转移到PANC-1细胞中，而不会导致明显的细胞死亡。这种非侵入性基因转移方法可能是未来胰腺癌安全临床基因治疗的有用工具。
Introduction: The purpose of this study was to determine whether ultrasound exposure combined with microbubble destruction could be used to enhance non-viral gene delivery in human pancreatic carcinoma cells (PANC-1).Methods: The study was performed with four experimental groups: Group P, plasmid alone; Group P+M, plasmid and microbubbles; Group P+U, plasmid and ultrasound; Group P+U+M, plasmid with ultrasound and microbubbles. Plasmid DNA encoding enhanced green fluorescent protein (pEGFP) was gently mixed with commercially available ultrasound microbubble contrast agents (SonoVue (R); Bracco Diagnostics Inc, Milan, Italy) in Group P+M and Group P+U+M. The different combinations of DNA and DNA plus microbubbles were added to cultured PANC-1 cells under different conditions. Transfection efficiency and cell viability were assessed by FACS analysis (Becton Dickinson, San Jose, CA, USA), confocal laser scanning microscopy, and trypan blue staining.Results: The results demonstrated that microbubbles with ultrasound exposure could significantly enhance the reporter gene expression as compared with other groups (Group P+U+M, 21.4%+/- 3.16%; Group P, 2.9%+/- 0.45%; Group P+M, 3.1%+/- 0.51%; Group P+U, 6.1%+/- 1.27%; P < 0.01). No statistically significant difference was observed in the PANC-1 cell viability between Group P+U+M and other groups (P > 0.05).Conclusions: Our in-vitro findings suggest that ultrasound-mediated microbubble destruction has the potential to promote efficient gene transfer into PANC-1 cells without significant cell death. This non-invasive gene transfer method may be a useful tool for safe clinical gene therapy of pancreatic cancer in the future.