Department of Thermobiology, Institute of Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236, Lodz, Poland.
Cell Biol Int. 2013 Dec;37(12):1330-9. doi: 10.1002/cbin.10165. Epub 2013 Sep 12.
We have investigated the mode of cell death induced by a new microtubule-stabilizing agent, epothilone B (EpoB, patupilone), and a clinically used medicine, paclitaxel (PTX), in normal ovarian cells. Using fluorescence microscopy, polyacrylamide gel electrophoresis preceding Western blot analysis, as well as spectrofluorimetric and colorimetric detection, we demonstrate that, compared to EpoB, PTX induced high time-dependent morphological and biochemical changes typical of apoptosis. Induction of apoptosis followed an early increase in p53 levels. Apoptosis reached its maximum at 24-48 h. At the same time, there was a significant increase in caspase-9 and -3 activity and PARP fragmentation, which suggests that an intrinsic path was involved. Apoptosis in MM14 cells was increased more by PTX than EpoB, and also induced more necrosis responsible for inflammation (1.4-fold) than EpoB.
我们研究了一种新型微管稳定剂——埃坡霉素 B（EpoB，紫杉醇）和一种临床用药——紫杉醇（PTX）在正常卵巢细胞中诱导细胞死亡的方式。通过荧光显微镜、聚丙烯酰胺凝胶电泳前的 Western blot 分析以及荧光光度法和比色法检测，我们证明与 EpoB 相比，PTX 诱导了典型的凋亡的高时间依赖性形态和生化变化。细胞凋亡伴随着 p53 水平的早期增加。凋亡在 24-48 小时达到最大值。同时，caspase-9 和 -3 的活性以及 PARP 片段化显著增加，这表明涉及内在途径。PTX 诱导的 MM14 细胞凋亡比 EpoB 多，并且比 EpoB 诱导更多的炎症性坏死（增加 1.4 倍）。
