DNA transfer and cell killing in epidermoid cells by diagnostic ultrasound activation of contrast agent gas bodies in vitro
DNA transfer and cell killing in epidermoid cells by diagnostic ultrasound activation of contrast agent gas bodies in vitro
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DOI：
10.1016/s0301-5629(02)00783-4
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发表时间：
2003-04-01
影响因子：
2.9
通讯作者：
Song, JM
中科院分区：
文献类型：
--
作者：
Miller, DL;Dou, CY;Song, JM
通过对比辅助的低功率诊断超声（US）检查单层细胞中的声孔和细胞杀伤的DNA转移。在37 ℃水浴中，用1.5-MHz扫描头以约1 mm/s的速度对具有表皮样细胞单层的培养室进行扫描。对于DNA转移试验，将编码绿色荧光蛋白（GFP）的质粒加入培养基中，2天后通过流式细胞术评估GFP表达。在单独的测试中，处理后立即测定细胞杀伤。假手术组的GFP阳性细胞计数为0.4%（0.7% SD），2.3 MPa下暴露于2% Optison（R）造影剂时为3.7%（1.2% SD）。假手术组的死亡细胞比例为3.4%（1.7% SD），暴露组为28.6%（6.3% SD）。这两种效应随着Optison（R）浓度的增加和峰值稀疏压力幅度的增加而增加。对比辅助诊断US具有用于基因转移的潜在治疗应用，但似乎存在与细胞杀伤的权衡。（电子邮件：douglm@umich.edu）（C）2003年世界医学超声生物学联合会。
DNA transfer by sonoporation and cell killing in monolayer cells were examined by contrast-aided low-power diagnostic ultrasound (US). Culture chambers with epidermoid cell monolayers were scanned at about 1 mm/s with a 1.5-MHz scan head aimed upward at the chamber in a 37degreesC water bath. For DNA transfer tests, plasmids coding for green fluorescent protein (GFP) were added to the medium, and GFP expression was assessed by flow cytometry after 2 days. In separate tests, cell killing was determined immediately after treatment. GFP-positive cell counts were 0.4% (0.7% SD) for shams and 3.7% (1.2% SD) of cells for exposure at 2.3 MPa with 2% Optison(R) contrast agent. The fraction of dead cells was 3.4% (1.7% SD) in shams and 28.6% (6.3% SD) in exposed chambers. Both effects increased for increasing Optison(R) concentration and increasing peak rarefactional pressure amplitude. Contrast-aided diagnostic US has a potential therapeutic application for gene transfer, but a trade-off appears to exist with cell killing. (E-mail: douglm@umich.edu) (C) 2003 World Federation for Ultrasound in Medicine Biology.