Shi Mingfang, Liu Bangzhong, Liu Guanghua, Wang Ping, Yang Mingzhen, Li Yun, Zhou Jian
Department of Rehabilitation, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Department of Rehabilitation, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Ultrasonics. 2016 Jan;64:43-53. doi: 10.1016/j.ultras.2015.07.011. Epub 2015 Jul 26.
The present study was conducted to determine whether low intensity-pulsed ultrasound (LIPUS) could induce apoptosis of human hepatocellular carcinoma cells, SMMC-7721, and to define the mechanism of ultrasound-induced apoptosis, in vitro. MTT assay was used to measure cell proliferation. Apoptosis was investigated by multiple methods such as flow cytometry, DNA fragmentation, Ca(2+) mobilizations, pro- and anti-apoptotic protein expression, and light as well as ultramicroscopic morphology. The results provide evidence that LIPUS induced a dose-dependent effect on cell viability and apoptosis of SMMC-7721 cells. Specifically, exposure of cells to >0.5 W/cm(2) intensity significantly increased cell apoptosis, caused shifts in cell cycle phase, and induced structural changes. Ultrasound significantly increased intracellular Ca(2+) concentrations and modulated expression of caspase-3, Bcl-2 and Bax. The findings suggest that this novel technology can be used to induce SMMC-7721 apoptosis via the Ca(2+)/mitochondrial pathway and could potentially be of clinical use for the treatment of hepatocellular carcinoma (SMMC-7721 cell line) and other cancers.
本研究旨在体外确定低强度脉冲超声（LIPUS）是否能诱导人肝癌细胞SMMC - 7721凋亡，并明确超声诱导凋亡的机制。采用MTT法检测细胞增殖。通过多种方法研究凋亡情况，如流式细胞术、DNA片段化、Ca(2+)动员、促凋亡和抗凋亡蛋白表达以及光镜和超微镜形态学。结果表明，LIPUS对SMMC - 7721细胞的活力和凋亡具有剂量依赖性影响。具体而言，细胞暴露于强度>0.5 W/cm(2)时，细胞凋亡显著增加，细胞周期阶段发生改变，并诱导结构变化。超声显著增加细胞内Ca(2+)浓度，并调节caspase - 3、Bcl - 2和Bax的表达。这些发现表明，这项新技术可通过Ca(2+)/线粒体途径诱导SMMC - 7721凋亡，可能在临床上用于治疗肝癌（SMMC - 7721细胞系）及其他癌症。
