Key Laboratory of Food Nutrition and Safety, Ministry of Education, College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China; QingYunTang Biotech Co., Ltd., No. 14, Zhonghe Street, Beijing Economic-Technological Development Area, Beijing 100176, China.
Key Laboratory of Food Nutrition and Safety, Ministry of Education, College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China.
Int J Biol Macromol. 2020 May 1;150:449-458. doi: 10.1016/j.ijbiomac.2020.02.010. Epub 2020 Feb 3.
In this study, a water-soluble polysaccharide was extracted from Polygonum tenuifolia to inhibit the proliferation of SPC-A-1 cells by the FAS/FAS-L-mediated pathway and autophagy. The molecular weight, composition, and structure of the Polygala tenuifolia polysaccharide (PTP) was detected by HPLC, HPAEC-PAD, NMR and FT-IR. The purified polysaccharide was composed of Ara, Gal, and Glc (molar ratio: 2.6:1.8:1.0) with α- and β-configurations. Morphological changes were observed with microscopes, and the cell apoptosis-related markers detected by flow cytometry indicated that apoptosis and autophagy occurred in the SPC-A-1 cells. Western blot analysis showed that the expression of proteins was related to apoptosis and autophagy. The death receptor pathway demonstrated the up-regulated expression of FAS, ligand FAS-L, and FADD, which led to a cascade reaction of the caspase family that induced cell apoptosis. The up-regulation of LC 3B-II and the down-regulation of P62 indicated the occurrence of autophagy. In summary, these results showed that PTP can induce FAS/FAS-L-mediated apoptosis and autophagy in SPC-A-1 cells, and provide a strong theoretical basis for tumor prevention and clinical application of PTP in the future.
在这项研究中，从虎杖中提取了一种水溶性多糖，通过 FAS/FAS-L 介导的途径和自噬来抑制 SPC-A-1 细胞的增殖。采用 HPLC、HPAEC-PAD、NMR 和 FT-IR 检测了虎杖多糖（PTP）的分子量、组成和结构。纯化的多糖由 Ara、Gal 和 Glc（摩尔比：2.6:1.8:1.0）组成，具有α-和β-构型。用显微镜观察形态变化，流式细胞术检测细胞凋亡相关标志物表明 SPC-A-1 细胞发生了凋亡和自噬。Western blot 分析表明，蛋白表达与细胞凋亡和自噬有关。死亡受体途径显示 FAS、配体 FAS-L 和 FADD 的表达上调，导致半胱氨酸蛋白酶家族的级联反应，诱导细胞凋亡。LC3B-II 的上调和 P62 的下调表明自噬的发生。综上所述，这些结果表明 PTP 可以诱导 SPC-A-1 细胞中 FAS/FAS-L 介导的凋亡和自噬，为 PTP 在未来肿瘤预防和临床应用提供了强有力的理论依据。
