Polyphenols Extracted from Enteromorpha clathrata Induce Apoptosis in Hepa1-6 Cell by Activating the Mitochondrial Apoptosis Signaling Pathways  

作　　者：ZHANG Panxue SUN Bolun TONG Jingjing XIA Geran ZHANG Jinjie LI Chao YANG Wenge 

机构地区：[1]College of Food and Pharmaceutical Sciences,Ningbo University,Ningbo 315211,China [2]Key Laboratory of Animal Protein Food Deep Processing Technology of Zhejiang Province,Ningbo University,Ningbo 315211,China

出　　处：《Journal of Ocean University of China》2023年第5期1393-1402,共10页中国海洋大学学报（英文版）

基　　金：the National Key R&D Program of China(No.2018YFD0901105);the Ningbo Natural Science Foundation(No.202003N4128);the Scientific Research Foundation of Graduate School of Ningbo University(No.IF2021085)。

摘　　要：Green alga Enteromorpha clathrata(E.clathrata)contains a variety of bioactive compounds,including polysaccharides,polyphenols and fat-soluble pigments etc.,among which polyphenols exhibit a wide range of medicinal properties.E.clathrata polyphenols(ECPs)have shown various biological activities such as antioxidant,anti-inflammatory and antidiabetic effects;however,the potential of ECPs as an anti-cancer reagent remains unclear.The aim of this study was to investigate the anti-tumor activity and underlying mechanisms of ECPs on hepatocellular carcinoma.The cytotoxicity of Hepa1-6 cells was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and lactate dehydrogenase(LDH)assay.Flow cytometry and fluorescence microscope analysis of cell apoptosis after annexin V-fluorescein isothiocyanate(FITC)/propidium iodide(PI)staining.2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)assay was used for intracellular reactive oxygen species(ROS)detection.caspase-9 activity was determined using cspase-9 colorimetric assay.Mitochondrial transmembrane potential(Δψm)was measured using JC-1.Western blot and quantitative real-time PCR(qPCR)were used to assess the expressions of the apoptosis regulators Bax,Bcl-2,cytochrome c and caspase-3.It was found that ECPs showed a dose-dependent cytotoxicity against Hepa1-6 cells by inducing apoptosis.The apoptosis in ECPs-treated Hepa1-6 cells was accompanied by the loss of mitochondrial membrane potential,elevated ROS generation,increased release of mitochondrial cytochrome c,and up-regulation of caspase-9 and caspase-3.The expressions of Bax(pro-apoptotic molecule)and Bcl-2(apoptosis suppressor)were up-regulated and down-regulated,respectively,at both mRNA and protein levels.These molecular alterations revealed that ECPs caused apoptosis of cells through the mitochondrial pathway,suggesting that ECPs are potential candidates to be developed for liver cancer treatment.

关 键 词：Enteromorpha clathrate POLYPHENOL anti-cancer activity apoptosis mitochondrial-dependent pathway 

分 类 号：Q949.9[生物学—植物学]