International Journal of Radiation Biology ( IF 2.1 ) Pub Date : 2015-02-06 , DOI: 10.3109/09553002.2015.1001532 Xinna Wang 1 , Jianmei Luo , Albert Wingnang Leung , Yajun Li , Hongwei Zhang , Chuanshan Xu
Purpose: To study subcellular localization of hypocrellin B in hepatocellular carcinoma cells, and hypocrellin B-mediated sonodynamic action-induced cell damage.
Materials and methods: After incubation with 2.5 μM of hypocrellin B, human hepatocellular carcinoma HepG2 cells were exposed to ultrasound waves for 8 sec at an intensity of 0.46 W/cm2. Clonogenic survival of HepG2 cells was measured using a colony forming assay and light microscope. Ultrastructural morphology was observed using transmission electron microscope (TEM) and mitochondrial membrane potential (MMP) was assessed using confocal laser scanning microcope (CLSM) after rhodamine 123 staining. Additionally, subcellular localization of hypocrellin B in HepG2 cells with organelle probe staining was also observed using CLSM.
Results: The colony forming units of HepG2 cells decreased substantially after sonodynamic treatment. The results of TEM showed microvilli disappearance, apoptotic body formation, swollen mitochondria with loss of cristae and mitochondrial myelin-like features (or membrane whorls). Collapse of MMP was found in the treated cells. Hypocrellin B was distributed in mitochondria and lysosomes as well as in endoplasmic reticulum and Golgi apparatus.
Conclusions: The findings demonstrated that sonodynamic action of hypocrellin B induced mitochondrial damage, survival inhibition, and apoptosis of HepG2 cells. Additionally, other subcellular organelles such as endoplasmic reticulum, Golgi apparatus and lysosomes were also the targets of hypocrellin B-mediated sonodynamic action as well as mitochondria.
中文翻译:
![](https://scdn.x-mol.com/jcss/images/paperTranslation.png)
肝癌细胞中的hypocrellin B:亚细胞定位和声动力学损伤。
目的:研究降钙素B在肝癌细胞中的亚细胞定位,以及降钙素B介导的声动力学作用诱导的细胞损伤。
材料和方法:与2.5μM的hypercrelinin B孵育后,将人类肝细胞癌HepG2细胞以0.46 W / cm 2的强度暴露于超声波中8秒钟。使用集落形成测定法和光学显微镜测量HepG2细胞的克隆存活。罗丹明123染色后,使用透射电子显微镜(TEM)观察超微结构形态,并使用共聚焦激光扫描显微技术(CLSM)评估线粒体膜电位(MMP)。另外,还使用CLSM观察到了用细胞器探针染色的HepG2细胞中的hypercrerinin B的亚细胞定位。
结果:经声动力学处理后,HepG2细胞的集落形成单位显着减少。透射电镜的结果显示微绒毛消失,凋亡小体形成,线粒体肿胀并失去ista和线粒体髓鞘样特征(或膜轮状)。在处理过的细胞中发现MMP崩溃。降钙素B分布在线粒体和溶酶体以及内质网和高尔基体中。
结论:研究结果表明,hypocrelinin B的声波动力学作用可诱导线粒体损伤,存活抑制和HepG2细胞凋亡。另外,其他亚细胞器,例如内质网,高尔基体和溶酶体,也是降钙素B介导的声波动力学作用以及线粒体的靶标。