半边旗二萜类成分PAG通过ROS诱导的线粒体凋亡克服肝癌多药耐药

Pteris semipinna-derived PAG reverses multidrug resistance through ROS-induced mitochondrial apoptosis in hepatocarcinoma cells

目的研究半边旗二萜类成分PAG对人肝癌耐药细胞株增殖与凋亡机制。方法 HepG2/ADM细胞用不同浓度（0～75μmol/L）PAG处理24、48、72 h,分别用MTT、流式细胞术、Western blot、DCFH-DA法检测细胞增殖、细胞周期、细胞凋亡、凋亡相关蛋白表达、活性氧自由基（ROS）水平。结果 PAG呈剂量依赖性抑制细胞增殖及caspase 3、PARP和Bcl-2表达,增加G2/M期细胞分布、细胞凋亡、细胞内ROS水平及actived-caspase 3、cleaved-PARP和Bax表达,促进线粒体中细胞色素C释放到胞质;1 mmol/L乙酰半胱氨酸可阻断PAG对细胞增殖的抑制作用。结论半边旗二萜类成分PAG可通过ROS诱导的线粒体凋亡克服肝癌多药耐药。

Objective To investigate the mechanism of Pteris semipinna-derived PAG on proliferation and apoptosis of human hepatocarcinoma cell line with multidrug resistance. Methods After treated with different concentrations of PAG （0-75 μmol/L）, proliferation, cycle, apoptosis, apoptosis-related protein expression and reactive oxygen species （ROS） level in HepG2/ADM cells were evaluated by MTT, flow cytometry, Western bolt and DCFH-DA, respectively. Results PAG inhibited dose-dependently cell proliferation and expression of caspase 3, PARP and Bcl-2, but increased G2/M distribution, apoptosis, cellular ROS level, cytochrome C release from mitochondria, and expression of actived-caspase 3, cleaved-PARP and Bax. Pretreatment with 1 mmol/L acetylcysteine abolished PAG-induced growth inhibition. Conclusion Pteris semipinna-derived PAG could reverse the multidrug resistance of hepatocarcinoma cells through ROS-induced mitochondrial apoptosis.