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低频超声联合微泡可改善脂质体介导的野生型P53 DNA转染人前列腺癌细胞的效果。

Liposome-mediated transfection of wild-type P53 DNA into human prostate cancer cells is improved by low-frequency ultrasound combined with microbubbles.

作者信息

Bai Wen-Kun, Zhang Wei, Hu Bing, Ying Tao

机构信息

Department of Ultrasound in Medicine, Shanghai Institute of Ultrasound in Medicine, Sixth People's Hospital Affiliated to Shanghai Jiao Tong University, Shanghai 200233, P.R. China.

出版信息

Oncol Lett. 2016 Jun;11(6):3829-3834. doi: 10.3892/ol.2016.4477. Epub 2016 Apr 20.

Abstract

Prostate cancer is a common type of cancer in elderly men. The aim of the present study was to evaluate the effects of ultrasound exposure in combination with SonoVue microbubbles on liposome-mediated transfection of wild-type P53 genes into human prostate cancer cells. PC-3 human prostate cancer cells were exposed to ultrasound; duty cycle was controlled at 20% (2 sec on, 8 sec off) for 5 min with and without SonoVue microbubble echo-contrast agent using a digital sonifier (frequency, 21 kHz; intensity, 46 mW/cm). The cells were divided into eight groups, as follows: Group A (SonoVue + wild-type P53), group B (ultrasound + wild-type P53), group C (SonoVue + ultrasound + wild-type P53), group D (liposome + wild-type P53), group E (liposome + SonoVue + wild-type P53), group F (liposome + wild-type P53 + ultrasound), group G (liposome + wild-type P53 + ultrasound + SonoVue) and the control group (wild-type P53). Following treatment, a hemocytometer was used to measure cell lysis, reverse transcription-quantitative polymerase chain reaction and western blotting were performed to detect P53 gene transfection efficiency, Cell Counting Kit-8 was employed to reveal cell proliferation and Annexin V/propidium iodide staining was used to determine cell apoptosis. Cell lysis was minimal in each group. Wild-type P53 gene and protein expression were significantly increased in the PC-3 cells in group G compared with the control and all other groups (P<0.01). Cell proliferation was significantly suppressed in group G compared with the control group and all other groups (P<0.01). Cell apoptosis levels in group G were significantly improved compared with the control group and all other groups (P<0.01). Thus, the results of the present study indicate that the use of low-frequency and low-energy ultrasound in combination with SonoVue microbubbles may be a potent physical method for increasing liposome gene delivery efficiency.

摘要

前列腺癌是老年男性中常见的一种癌症。本研究的目的是评估超声暴露联合声诺维微泡对脂质体介导的野生型P53基因转染人前列腺癌细胞的影响。将PC-3人前列腺癌细胞暴露于超声下;使用数字超声仪(频率21kHz;强度46mW/cm²),在有和声诺维微泡回声造影剂的情况下,将占空比控制在20%(开2秒,关8秒)持续5分钟。细胞分为八组,如下:A组(声诺维+野生型P53),B组(超声+野生型P53),C组(声诺维+超声+野生型P53),D组(脂质体+野生型P53),E组(脂质体+声诺维+野生型P53),F组(脂质体+野生型P53+超声),G组(脂质体+野生型P53+超声+声诺维)和对照组(野生型P53)。处理后,使用血细胞计数器测量细胞裂解情况,进行逆转录-定量聚合酶链反应和蛋白质印迹法检测P53基因转染效率,使用细胞计数试剂盒-8检测细胞增殖情况,并用膜联蛋白V/碘化丙啶染色法测定细胞凋亡情况。每组细胞裂解程度均最小。与对照组及所有其他组相比,G组PC-3细胞中野生型P53基因和蛋白表达显著增加(P<0.01)。与对照组及所有其他组相比,G组细胞增殖受到显著抑制(P<0.01)。与对照组及所有其他组相比,G组细胞凋亡水平显著改善(P<0.01)。因此,本研究结果表明,低频低能量超声联合声诺维微泡可能是提高脂质体基因递送效率的一种有效物理方法。

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