SONOPORATION AS A CELLULAR STRESS: INDUCTION OF MORPHOLOGICAL REPRESSION AND DEVELOPMENTAL DELAYS
SONOPORATION AS A CELLULAR STRESS: INDUCTION OF MORPHOLOGICAL REPRESSION AND DEVELOPMENTAL DELAYS
复制标题
DOI：
10.1016/j.ultrasmedbio.2013.01.008
复制
发表时间：
2013-06-01
影响因子：
2.9
通讯作者：
Yu, Alfred C. H.
中科院分区：
文献类型：
--
作者：
Chen, Xian;Wan, Jennifer M. F.;Yu, Alfred C. H.
声孔作为一种药物/基因传递的范例建立，它是至关重要的，以考虑这种膜渗透策略对活细胞的生物学影响。在这里，我们提供了新的见解声穿孔细胞的影响，通过证明在体外，这种方式的渗透质膜可能会无意中诱导抑制细胞功能，即使在增强外源性分子的摄取。悬浮型（HL-60）和单层（ZR-75-30）细胞被认为是在这项调查中，他们经常暴露于1-MHz脉冲超声（脉冲长度，100个周期;脉冲重复频率，1 kHz;曝光周期，60 s）（空间平均峰值负压，0.45 MPa）和在微泡存在下（细胞：泡比，10：1）。使用共聚焦显微镜检查声穿孔细胞（鉴定为具有钙黄绿素内化的细胞）的暴露后形态，并使用流式细胞术分析其细胞周期进展动力学。结果表明，对于两种类型的细胞研究，sonoporated细胞表现出膜收缩和细胞内脂质积累超过2小时的时间。此外，与正常细胞相比，声穿孔细胞的脱氧核糖核酸合成持续时间显着延长，表明细胞周期进展延迟。已知这些特征是细胞应激反应的特征，表明声致穿孔确实构成对活细胞的应激。这个问题可能需要在优化声致穿孔用于药物/基因递送目的时解决。另一方面，它增加了通过声孔作用开发其他治疗应用的机会。（E-mail：alfred. hku.hk）（C）2013年世界医学和生物学超声联合会。
For sonoporation to be established as a drug/gene delivery paradigm, it is essential to account for the biological impact of this membrane permeation strategy on living cells. Here we provide new insight into the cellular impact of sonoporation by demonstrating in vitro that this way of permeating the plasma membrane may inadvertently induce repressive cellular features even while enhancing exogenous molecule uptake. Both suspension-type (HL-60) and monolayer (ZR-75-30) cells were considered in this investigation, and they were routinely exposed to 1-MHz pulsed ultrasound (pulse length, 100 cycles; pulse repetition frequency, 1 kHz; exposure period, 60 s) with calibrated field profile (spatial-averaged peak negative pressure, 0.45 MPa) and in the presence of microbubbles (cell: bubble ratio, 10: 1). The post-exposure morphology of sonoporated cells (identified as those with calcein internalization) was examined using confocal microscopy, and their cell cycle progression kinetics were analyzed using flow cytometry. Results show that for both cell types investigated, sonoporated cells exhibited membrane shrinkage and intra-cellular lipid accumulation over a 2-h period. Also, as compared with normal cells, the deoxyribonucleic acid synthesis duration of sonoporated cells was significantly lengthened, indicative of a delay in cell cycle progression. These features are known to be characteristics of a cellular stress response, suggesting that sonoporation indeed constitutes as a stress to living cells. This issue may need to be addressed in optimizing sonoporation for drug/gene delivery purposes. On the other hand, it raises opportunities for developing other therapeutic applications via sonoporation. (E-mail: alfred.yu@hku.hk) (C) 2013 World Federation for Ultrasound in Medicine & Biology.