State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR China.
State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, PR China.
Int J Biol Macromol. 2019 Mar 15;125:1232-1241. doi: 10.1016/j.ijbiomac.2018.09.098. Epub 2018 Sep 17.
Grifola frondosa is a widely eaten and medicinal fungus. In this study, we extracted a cold-water-soluble polysaccharide from Grifola frondosa (cGFP) and investigated its effects on the proliferation and apoptosis of human hepatoma HepG2 cells. MTT assay showed that cGFP induced apoptosis of HepG2 cells in a dose-dependent manner. Flow cytometry analysis showed that cGFP induced apoptosis in HepG2 cells through S phase arrest. The distribution of cells at different apoptotic stages was determined by Annexin V-FITC and Propidium Iodide (PI) staining. Scanning electron microscopy (SEM) results indicated that cGFP induced typical apoptotic morphological features in HepG2. Mitochondrial membrane potential was reduced according to the screening of JC-1 staining. And western blot analysis of Bax, Bcl-2, cytochrome C (Cyto-c), caspase-3, and caspase-9 further demonstrated that the cGFP-induced apoptosis effect functioned through the mitochondrial pathway. Further analysis by qRT-PCR showed that Bax expression increased and Bcl-2 expression decreased. These findings suggested that cGFP could inhibit the proliferation of HepG2 cells and induce apoptosis mainly through the intrinsic activation mitochondrial pathway.
灰树花是一种广泛食用和药用的真菌。在本研究中，我们从灰树花（cGFP）中提取了一种冷水溶性多糖，并研究了其对人肝癌 HepG2 细胞增殖和凋亡的影响。MTT 试验表明，cGFP 呈剂量依赖性诱导 HepG2 细胞凋亡。流式细胞术分析表明，cGFP 通过 S 期阻滞诱导 HepG2 细胞凋亡。通过 Annexin V-FITC 和碘化丙啶（PI）染色确定不同凋亡阶段细胞的分布。扫描电子显微镜（SEM）结果表明，cGFP 诱导 HepG2 出现典型的凋亡形态特征。根据 JC-1 染色筛选，线粒体膜电位降低。Bax、Bcl-2、细胞色素 C（Cyto-c）、caspase-3 和 caspase-9 的 Western blot 分析进一步表明，cGFP 诱导的细胞凋亡作用是通过线粒体途径发挥的。qRT-PCR 进一步分析表明，Bax 表达增加，Bcl-2 表达减少。这些结果表明，cGFP 可以抑制 HepG2 细胞的增殖，并主要通过内在激活的线粒体途径诱导细胞凋亡。
