Wang Ping, Leung Albert Wingnang, Xu Chuanshan
Department of Ultrasound Medicine, The Affiliated Hospital of North Sichuan Medical College, Sichuan;
Exp Ther Med. 2011 Sep;2(5):849-852. doi: 10.3892/etm.2011.317. Epub 2011 Jul 1.
Ultrasound therapy, as a non-invasive modality, has been attracting extensive attention in the management of malignant tumors. The present study aimed to investigate low-intensity ultrasound-induced cellular destruction and autophagy in nasopharyngeal carcinoma cells in vitro. Nasopharyngeal carcinoma CNE2 cells were subjected to ultrasound exposure, as tumor model cells, at an intensity of 1.35 W/cm(2). Cytotoxicity was investigated 24 h after ultrasound treatment. Nuclear damage was observed using nuclear staining with Hoechst 33258. Mitochondrial dysfunction was measured using confocal laser scanning microscopy with rhodamine123 staining. Mitochondrial morphology and autophagy were observed using transmission electron microscopy (TEM). Low-intensity ultrasound significantly killed CNE2 cells proportional to the ultrasonic treatment time. Upon nuclear staining, nuclear condensation and typical apoptotic bodies were noted in the CNE2 cells exposed to ultrasound wave for 12 sec. A collapse in mitochondrial membrane potential was noted in the treated cells. Upon TEM, swollen mitochondria, more vacuoles and autophagy were noted after ultrasound treatment. Our findings demonstrate that low-intensity ultrasound significantly damages CNE2 cells and emphasize that autophagy may be an important event in ultrasound-induced cell death.
超声治疗作为一种非侵入性治疗方式,在恶性肿瘤的治疗中已引起广泛关注。本研究旨在体外研究低强度超声诱导的鼻咽癌细胞的细胞破坏和自噬。将鼻咽癌细胞系CNE2细胞作为肿瘤模型细胞,以1.35 W/cm(2)的强度进行超声照射。超声处理24小时后研究细胞毒性。使用Hoechst 33258进行核染色观察核损伤。使用罗丹明123染色的共聚焦激光扫描显微镜测量线粒体功能障碍。使用透射电子显微镜(TEM)观察线粒体形态和自噬。低强度超声显著杀死CNE2细胞,其与超声处理时间成正比。经核染色后,在暴露于超声波12秒的CNE2细胞中观察到核浓缩和典型的凋亡小体。在处理过的细胞中观察到线粒体膜电位的崩溃。经TEM观察,超声处理后可见线粒体肿胀、更多空泡和自噬现象。我们的研究结果表明,低强度超声显著损伤CNE2细胞,并强调自噬可能是超声诱导细胞死亡中的一个重要事件。