State Key Laboratory of Food Nutrition and Safety, Ministry of Education, College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, No. 29, 13th Street, TEDA, 300457, Tianjin, People's Republic of China.
QingYunTang Biotech (Beijing) Co., Ltd, Beijing Economic-Technological Development Area, No. 14, Zhonghe Street, 100176, Beijing, China.
Glycoconj J. 2020 Aug;37(4):413-422. doi: 10.1007/s10719-020-09932-0. Epub 2020 Jun 15.
In this study, a novel water soluble polysaccharide (named GFP-4) was extracted from Grifola frondosa at 4 C, and its preliminary structure and inhibitory effects on human gastric carcinoma MKN-45 cells through the Fas/FasL death receptor apoptosis pathway were investigated. High-performance gel permeation chromatography (HPGPC), fourier-transform infrared spectroscopy (FT-IR), and ion chromatography (IC) results showed that GFP-4 was a 1.09 × 10 Da neutral hetero polysaccharide with pyranose rings, and α- and β-type glycosidic linkages that contained galactose, glucose, and mannose at a molar ratio of 1.00:3.45:1.19. MTT results indicated that GFP-4 significantly inhibited the proliferation of MKN-45 cells in a concentration-dependent manner. The H&E staining and Hoechst 33342/PI double staining results showed that GFP-4-treated MKN-45 cells were subjected to underwent typical apoptotic morphologic changes such as nuclear pyknosis, chromatin condensation, and an increase of membrane permeability. Annexin V-FITC/PI double staining, cell cycle analysis, and western blot results revealed the GFP-4 induced MKN-45 cells apoptosis through the Fas/FasL-mediated death receptor pathway with cells arrested at the G0/G1 phase. These data indicate that GFP-4 is a promising candidate for treating gastric cancer and provide a theoretical basis for the future development and utilization of G. frondosa clinically.
在这项研究中，从灰树花中在 4°C 下提取了一种新型水溶性多糖（命名为 GFP-4），并通过 Fas/FasL 死亡受体凋亡途径研究了其对人胃癌 MKN-45 细胞的初步结构和抑制作用。高效凝胶渗透色谱（HPGPC）、傅里叶变换红外光谱（FT-IR）和离子色谱（IC）结果表明，GFP-4 是一种 1.09×10 Da 的中性杂多糖，具有吡喃糖环和α-和β-型糖苷键，含有半乳糖、葡萄糖和甘露糖，摩尔比为 1.00:3.45:1.19。MTT 结果表明，GFP-4 以浓度依赖的方式显著抑制 MKN-45 细胞的增殖。H&E 染色和 Hoechst 33342/PI 双重染色结果表明，GFP-4 处理的 MKN-45 细胞经历了典型的凋亡形态变化，如核固缩、染色质浓缩和膜通透性增加。Annexin V-FITC/PI 双重染色、细胞周期分析和 Western blot 结果表明，GFP-4 通过 Fas/FasL 介导的死亡受体途径诱导 MKN-45 细胞凋亡，细胞停滞在 G0/G1 期。这些数据表明 GFP-4 是治疗胃癌的有前途的候选药物，为灰树花在临床上的未来开发和利用提供了理论依据。
