Hispolon Induces Apoptosis and Cell Cycle Arrest of Human Hepatocellular Carcinoma Hep3B Cells by Modulating ERK Phosphorylation
Hispolon Induces Apoptosis and Cell Cycle Arrest of Human Hepatocellular Carcinoma Hep3B Cells by Modulating ERK Phosphorylation
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DOI：
10.1021/jf201289e
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发表时间：
2011-07-13
影响因子：
6.1
通讯作者：
Hu, Miao-Lin
中科院分区：
文献类型：
--
作者：
Huang, Guan-Jhong;Deng, Jeng-Shyan;Hu, Miao-Lin
Hispolon是Phellinus igniarius中的一种活性酚类化合物，Phellinus igniarius是一种最近被证明具有抗氧化、抗炎和抗癌活性的蘑菇。本研究采用MTT法、DNA片段化、DAPI（4,6-二氨基-2-苯基吲哚二盐酸）染色、流式细胞术等方法研究组蛋白蛋白对人肝癌Hep3B细胞的抗增殖作用。Hispolon抑制Hep3B细胞的生长具有时间依赖性和剂量依赖性，通过流式细胞术分析和凋亡细胞死亡诱导S期细胞周期阻滞，如DNA阶梯所示。hispolon诱导的s期阻滞与细胞周期蛋白a、E和细胞周期蛋白依赖性激酶（CDK） 2的蛋白表达显著降低相关，同时诱导p21waf1/Cip1和p27Kip1。Hep3B细胞暴露于组蛋白蛋白会导致凋亡，这可以通过caspase激活、PARP切割和DNA断裂来证明。Hispolon处理也激活了JNK、p38 MAPK和ERK的表达。ERK （PB98095）抑制剂，而JNK （SP600125）和p38 MAPK （SB203580）抑制剂，抑制了hispolon诱导的Hep3B细胞的s期阻滞和凋亡。这些发现建立了MARK通路与组蛋白蛋白诱导的Hep3B细胞周期阻滞和凋亡之间的机制联系。
Hispolon is an active phenolic compound of Phellinus igniarius, a mushroom that has recently been shown to have antioxidant, anti-inflammatory, and anticancer activities. This study investigated the antiproliferative effect of hispolon on human hepatocellular carcinoma Hep3B cells by using the MTT assay, DNA fragmentation, DAPI (4,6-diamidino-2-phenylindole dihydrochloride) staining, and flow cytometric analyses. Hispolon inhibited cellular growth of Hep3B cells in a time-dependent and dose-dependent manner, through the induction of cell cycle arrest at S phase measured using flow cytometric analysis and apoptotic cell death, as demonstrated by DNA laddering. Hispolon-induced S-phase arrest was associated with a marked decrease in the protein expression of cyclins A and E and cyclin-dependent kinase (CDK) 2, with concomitant induction of p21waf1/Cip1 and p27Kip1. Exposure of Hep3B cells to hispolon resulted in apoptosis as evidenced by caspase activation, PARP cleavage, and DNA fragmentation. Hispolon treatment also activated JNK, p38 MAPK, and ERK expression. Inhibitors of ERK (PB98095), but not those of JNK (SP600125) and p38 MAPK (SB203580), suppressed hispolon-induced S-phase arrest and apoptosis in Hep3B cells. These findings establish a mechanistic link between the MARK pathway and hispolon-induced cell cycle arrest and apoptosis in Hep3B cells.