Ultrasonic enhancement of gene transfection in murine melanoma tumors
Ultrasonic enhancement of gene transfection in murine melanoma tumors
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DOI：
10.1016/s0301-5629(99)00105-2
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发表时间：
1999-11-01
影响因子：
2.9
通讯作者：
Thrall, BD
中科院分区：
文献类型：
--
作者：
Miller, DL;Bao, SP;Thrall, BD
采用B16小鼠黑色素瘤模型，在体内外评价超声对基因转导的促进作用。有教养的。将细胞接种于C57BL/6雌性小鼠皮下或体外培养10-14天后，用绿色荧光蛋白报告质粒(GFP)评价转基因效率，并与荧光素酶报告质粒(GFP)进行比较。US由类似Dornier HM-3型碎石机的系统提供，该系统在焦点处产生24.4 Mpa峰值正压和5.2 Mpa峰值负压幅度的冲击波(Sw)。将其与悬浮液混合，获得20亩L毫升(-1)，或瘤内注射，每毫升肿瘤提供0.2mgDNA。在体外1.2毫升的曝气室中保持0.2毫升的空气和在体内注入10%的空气可以促进声空化。在体外，800短波照射后细胞数下降到SHAMS的5.3%，培养2天后有1.4%的细胞表达GFP。在体内，400Sw照射后2天，切除肿瘤的活细胞回收率降至假手术组的4.2%，细胞转染率增加约8倍，达到细胞计数的2.5%(t检验p<；0.005)，这些结果表明，超声冲击波治疗诱导的强大肿瘤消融与基因转染率的提高是同步进行的。(C)1999年世界医学和生物学超声联合会。
The enhancement of gene transfection by ultrasound (US) was evaluated in vitro and in vivo using the B16 mouse melanoma model. Cultured. cells were either exposed in suspensions in vitro or implanted subcutaneously in female C57BL/6 mice for 10-14 days and, subsequently exposed, in vivo, For comparison to results with a luciferase plasmid, a reporter plasmid for green fluorescent protein (GFP) was used to evaluate transfection efficiency. US was supplied by a system, similar to a Dornier HM-3 lithotripter, that produced shock waves (SW) of 24.4 MPa peak positive and 5.2 MPa peak negative pressure amplitudes at the focus. The plasmids were mixed with the suspensions to achieve 20 mu L mL(-1), or were injected intratumorally to provide 0.2 mg DNA per mt of tumor. Acoustic cavitation was promoted by retaining 0.2 mL of air in the 1.2-mL exposure chambers in vitro and by injecting air at 10% of tumor volume in vivo, In vitro, cell counts declined to 5.3% of shams after 800 SW exposure, with 1.4% of the cells expressing GFP after 2 days of culture. In vivo, 2 days after 400 SW exposure, viable-cell recovery from excised tumors was reduced to 4.2% of shams and cell transfection was enhanced by a factor of about 8, reaching 2.5% of cell counts (p < 0.005 in t-test), These results show that strong tumor ablation induced by US shock wave treatment can be coupled with simultaneous enhancement of gene transfection. (C) 1999 World Federation for Ultrasound in Medicine & Biology.