摘要
目的:从基因及蛋白水平探讨连翘根醇提物(ethanol extract of Forsythia suspensa root,FSEER)诱导人食管癌TE-13细胞凋亡的作用机制。方法:不同质量浓度的FSEER(0.4、0.5和0.6mg/mL)作用于TE-13细胞24h后,采用FCM检测线粒体膜电位的变化;蛋白质印迹法检测细胞凋亡相关蛋白[caspase-3、caspase-8、caspase-9和细胞质中细胞色素C(cytochrome-C,Cyt-C)]经FSEER处理后在TE-13细胞中的表达情况;同时通过RT-PCR检测细胞凋亡相关基因Bax、Bad、Noxa、Bcl-2、Bcl-xl和Mcl-1mRNA的表达情况。结果:TE-13细胞经FSEER(0.4、0.5和0.6mg/mL)作用24h后,随FSEER作用浓度的升高,发生线粒体膜电位损伤的细胞数目逐渐增多,且明显多于对照组,差异有统计学意义(P<0.05)。蛋白质印迹法检测结果显示,经FSEER处理后,TE-13细胞中cleaved caspase-3和cleaved caspase-9的表达水平及细胞质中Cyt-C的表达水平均逐渐增加,与对照组相比,差异有统计学意义(P<0.05);而caspase-8的表达水平则无明显变化,与对照组相比,差异无统计学意义。RT-PCR检测结果显示,FSEER可以上调TE-13细胞中Bax、Bad和NoxamRNA的表达水平,下调Bcl-2、Bcl-xl和Mcl-1mRNA的表达水平。结论:FSEER可能是通过依赖于线粒体的细胞凋亡内源途径而诱导人食管癌TE-13细胞发生凋亡的。
Objective： To investigate the probable mechanism of FSEER （ethanol extract of Forsythia suspensa root）-induced apoptosis of esophageal cancer TE-13 cells in vitro. Methods： Change of mitochondrial membrane potential in TE-13 celsl after treatment with different concentrations of FSEER （0.4, 0.5 and 0.6 mg/mL） for 24 h was examined by FCM （flow cytometry）. The expressions of apoptosis-associated proteins [cleaved caspase-3, caspase-8, cleaved caspase-9 and Cyt-C （cytochrome-C）] in esophageal cancer cells after FSEER treatment were detected by Western blotting. The expression levels of Bax, Bad, Noxa, Bcl-2, Bcl-xl and Mcl-1 mRNAs in TE-13 cells after treatment with different concentrations of FSEER for 24 h were detected by RT （reverse transcription）-PCR. Results： FSEER （0.4, 0.5 and 0.6 mg/mL for 24 h） can significantly reduce the mitochondrial membrane potential of TE-13 cells in a dose-dependent manner, as compared with the TE-13 cells without any treatment （P 〈 0.05）. The result of Western blotting showed that the expression levels of cleaved caspase-3, cleaved-caspase-9 and Cyt-C in TE-13 cells after FSEER treatment were significantly increased but the expression level of caspase-8 had no change as compared with those of the TE-13 cells without any treatment （P 〈 0.05）. RT-PCR showed that FSEER could up-regulate the expression levels of Bax, Bad and Noxa mRNAs and down-regulate the expression levels of Bcl-2, Bcl-xl and McI-1 mRNAs. Conclusion： The underlying mechanism of FSEER-induced apoptosis of TE-13 cells may be related to the mitochondria-dependent pathway.
出处
《肿瘤》
CAS
CSCD
北大核心
2013年第3期239-244,共6页
Tumor
基金
河北省中医药管理局科研计划资助项目(编号:2011011)
