β-岚香酮酸对肝细胞癌细胞增殖和凋亡的影响及其机制
Effect of β-elemonic Acid on Proliferation and Apoptosis of Hepatocellular Car-cinoma Cells and Its Mechanism
邹晨 1乔凤杰 1方淼 2高亚婷 2马言璐 1高月求 2张鑫2
作者信息
- 1. 上海中医药大学附属曙光医院细胞免疫实验室,上海市中医临床重点实验室,上海,201203
- 2. 上海中医药大学附属曙光医院细胞免疫实验室,上海市中医临床重点实验室,上海,201203;上海中医药大学附属曙光医院肝病科,上海,201203
- 折叠
摘要
本研究旨在探讨β-岚香酮酸(β-elemonic acid,β-EA)对肝细胞癌(hepatocellular carcinoma,HCC)细胞增殖和凋亡的影响及其作用机制.利用CCK8法检测β-EA对人HCC细胞Huh7和PLC/PRF/5活力的影响;利用Annexin V-FTTC/7AAD染色检测细胞凋亡变化;利用流式细胞周期检测β-岚香酮酸对HCC细胞周期的影响;利用Western blot检测凋亡相关蛋白Bcl2、BAX、cleaved-caspase 3和细胞周期相关蛋白CDK1、Cyclin B1的表达.结合网络药理学预测β-EA抑制HCC细胞增殖的分子机制,检测PI3K/AKT信号通路蛋白的表达.通过构建H22皮下移植瘤模型,体内证实了 β-EA对HCC的生长抑制作用.结果表明,β-EA能够显著抑制Huh7和PLC/PRF/5细胞活力,诱导HCC细胞凋亡和G2/M期细胞阻滞,上调促凋亡蛋白cleaved-caspase 3、BAX,下调抗凋亡蛋白Bc12、细胞周期蛋白Cyclin B1、细胞周期依赖性激酶CDK1、p-PI3K、p-AKT的表达.皮下移植瘤模型证实β-EA能够有效降低瘤重、瘤体积和Ki67的表达.本研究结果表明β-EA能够通过PI3K/AKT信号通路诱导HCC细胞凋亡和G2/M期细胞阻滞.
Abstract
The aim of this study was to investigate the effect of β-elemonic acid(β-EA)on hepatocellular carcinoma(HCC)prolifera-tion and apoptosis and its mechanism.CCK8 assay was used to detect the effects of β-EA on the cell viability of human HCC cells Huh7 and PLC/PRF/5.Annexin V-FTTC/7AAD staining was used to detect cell apoptosis.The effect of β-EA on cell cycle of HCC cells was detected by flow cytometry.The expressions of apoptosis-related proteins Bc12,BAX,cleaved-caspase 3 and cell cycle related pro-teins CDK1 and Cyclin B1 were detected by Western blot.The molecular mechanism of β-EA inhibiting the proliferation of HCC cells was predicted by network pharmacology,and the protein expression of PI3K/AKT signaling pathway was detected.The growth inhibi-tion effect of β-EA on HCC was confirmed by constructing H22 subcutaneous transplantation tumor model in vivo.The results showed that β-EA significantly inhibited Huh7 and PLC/PRF/5 cell viability,induced cell apoptosis and G2/M cell arrest.The expression of pro-apoptotic protein cleaved-caspase 3 and BAX was up-regulated,and the expression of anti-apoptotic protein Bc12,cell cycle protein Cyclin B1,cycle-dependent kinases CDK1,p-PI3K,and p-AKT was down-regulated.The subcutaneous tumor transplantation model confirmed that β-EA can effectively reduce tumor weight,tumor volume,and Ki67 expression.The results of this study suggest that β-EA can induce apoptosis and G2/M cell arrest of HCC cells through PI3K/AKT signaling pathway.
关键词
肝细胞癌/β-岚香酮酸/细胞增殖/细胞凋亡/细胞周期阻滞Key words
Hepatocellular carcinoma/β-elemonic acid/Cell proliferation/Cell apoptosis/Cell cycle arrest引用本文复制引用
基金项目
国家自然科学基金(81973773)
上海中医药大学"杏林百人"人才项目()
上海市临床重点专科建设项目(中医专业)(shslczdzk01201)
上海市名老中医学术经验研究工作室项目(SHGZS-202246)
中医药领军人才项目岐黄学者支持项目()
出版年
2024
NETL
NSTL
万方数据