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丁香酚诱导癌相关成纤维细胞凋亡的研究
李彦美1, 马 静2, 张志伟,等2
1.西北农林科技大学 创新实验学院;2.中国科学院 生物物理研究所
摘要:
【目的】研究丁香酚诱导癌相关成纤维细胞(CAF)的凋亡表现及其相关机制,为丁香酚的开发利用提供参考。【方法】用0(CK),30,60,90,120 μmol/L丁香酚处理正常成纤维细胞(NF)24 h后,用MTT法检测细胞活力。用0(CK),30,60,90,120,150,180 μmol/L丁香酚处理CAF细胞24 h后,用MTT法检测细胞活力。用0(CK),30,60 μmol/L丁香酚处理CAF细胞24 h,用Annexin V FITC和PI双染色法对细胞染色后进行流式细胞分析。用60 μmol/L丁香酚处理CAF细胞0(CK),15,30,60,120 min,用Western blotting法检测凋亡相关蛋白表达水平的变化。【结果】30~120 μmol/L丁香酚处理对NF细胞活力无显著影响;30~180 μmol/L丁香酚处理对CAF细胞活力有显著或极显著的抑制作用,丁香酚对CAF细胞的半数抑制浓度为84.47 μmol/L。60 μmol/L丁香酚能够显著诱导CAF细胞凋亡。凋亡相关蛋白Bax、Bcl-2、P-P53、Caspase 9、Cleaved Caspase 3、Cytochrome C参与了丁香酚诱导的CAF细胞凋亡调控过程。【结论】丁香酚对CAF细胞有抑制作用,其作用可能是通过P53蛋白途径和内源性凋亡途径实现的。
关键词:  丁香酚  癌相关成纤维细胞  细胞凋亡  抗癌机制
DOI:
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基金项目:国家自然科学基金项目(31370910)
Eugenol induced apoptosis of cancer associated fibroblasts and its mechanism
LI Yanmei,MA Jing,ZHANG Zhiwei,et al
Abstract:
【Objective】The aim of this research was to study the effect and related mechanism of eugenol on cancer associated fibroblasts(CAF),so as to provide reference for its development and utilization.【Method】Normal fibroblasts were treated with eugenol at concentrations of 0 (CK),30,60,90,and 120 μmol/L for 24 h,and cell viability was detected by MTT assay.CAF cells were treated with eugenol at concentrations of 0 (CK),30,60,90,120,150,and 180 μmol/L for 24 h,and cell viability was determined by MTT assay.CAF cells were treated with eugenol at concentrations of 0 (CK),30,and 60 μmol/L for 24 h,and stained with Annexin V FITC and PI for flow cytometry.CAF cells were treated with 60 μmol/L eugenol for 0 (CK),15,30,60,and 120 min,and the expression levels of apoptosis related proteins were detected by western blotting.【Result】Treatment with 30-120 μmol/L eugenol had no significant effect on viability of NF cells,while treatment with 30-180 μmol/L eugenol had significant or very significant inhibitory effect on CAF cell viability.The median inhibitory concentration of eugenol on CAF cells was 84.47 μmol/L.Eugenol at 60 μmol/L significantly induced apoptosis of CAF cells.Apoptosis-related proteins including Bax,Bcl-2,P-P53,Caspase 9,Cleaved Caspase 3 and Cytochrome C were involved in eugenol induced apoptosis regulation of CAF cells.【Conclusion】Eugenol could inhibit the viability of cancer associated fibroblasts by promoting apoptosis through the P53 protein pathway and endogenous pathway.
Key words:  eugenol  cancer associated fibroblasts  cell apoptosis  antitumor mechanism

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