Department of Hepatobiliary Surgery, Drum Tower Clinical Medical College of Nanjing Medical University, Nanjing, Jiangsu 210008, China.
Department of Hepatobiliary Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, Jiangsu 210008, China.
Biomed Res Int. 2014;2014:682197. doi: 10.1155/2014/682197. Epub 2014 Jul 6.
Rosin, the traditional Chinese medicine, is reported to be able to inhibit skin cancer cell lines. In this report, we investigate the inhibitory effect against HCC cells of QC2, the derivative of rosin's main components dehydroabietic acid.
MTT assay was used to determine the cytotoxicity of QC2. Morphological changes were observed by time-lapse microscopy and transmission electron microscopy and the cytoskeleton changes were observed by laser-scanning confocal microscopy. Cytomembrane integrity and organelles damage were confirmed by detection of the reactive oxygen (ROS), lactate dehydrogenase (LDH), and mitochondrial membrane potential (Δψm). The underlying mechanism was manifested by Western blotting. The oncotic cell death was further confirmed by detection of oncosis related protein calpain.
Swelling cell type and destroyed cytoskeleton were observed in QC2-treated HCC cells. Organelle damage was visualized by transmission electron microscopy. The detection of ROS accumulation, increased LDH release, and decreased ATP and Δψm confirmed the cell death. The oncotic related protein calpain was found to increase time-dependently in QC2-treated HCC cells, while its inhibitor PD150606 attenuated the cytotoxicity.
Dehydroabietic acid derivative QC2 activated oncosis related protein calpain to induce the damage of cytomembrane and organelles which finally lead to oncosis in HCC cells.
中药松香据称能够抑制皮肤癌细胞系。在本报告中，我们研究了松香主要成分去氢枞酸的衍生物 QC2 对 HCC 细胞的抑制作用。
MTT 法测定 QC2 的细胞毒性。通过时差显微镜和透射电子显微镜观察形态变化，通过激光共聚焦扫描显微镜观察细胞骨架变化。通过检测活性氧（ROS）、乳酸脱氢酶（LDH）和线粒体膜电位（Δψm）来确认细胞质膜完整性和细胞器损伤。通过 Western blot 显示潜在机制。通过检测与胀亡相关的蛋白钙蛋白酶进一步证实胀亡性细胞死亡。
在 QC2 处理的 HCC 细胞中观察到肿胀细胞类型和破坏的细胞骨架。透射电子显微镜观察到细胞器损伤。ROS 积累、LDH 释放增加以及 ATP 和 Δψm 减少的检测证实了细胞死亡。在 QC2 处理的 HCC 细胞中，发现与胀亡相关的蛋白钙蛋白酶随时间增加，而其抑制剂 PD150606 则减弱了细胞毒性。
去氢枞酸衍生物 QC2 激活与胀亡相关的蛋白钙蛋白酶，导致 HCC 细胞的细胞质膜和细胞器损伤，最终导致胀亡。
