绿原酸通过下调Notch1的表达抑制食管癌细胞的克隆形成和侵袭转移

Chlorogenic acid suppresses colony formation and invasion of esophageal squamous cell carcinoma via down-regulating the expression of Notch1

目的考察Notch1在绿原酸(CGA)抑制食管癌过程中的作用及机制。方法采用表达谱芯片检测食管癌小鼠食管组织中的差异基因表达,并用免疫组化(IHC)染色方法进行验证;用CGA对不同的食管癌细胞系进行处理,通过Western blot检测不同细胞中Notch1的表达变化情况。采用RNAi技术敲降食管癌细胞中Notch1的表达,再加入CGA处理,利用克隆形成实验和Transwell实验检测食管癌细胞的克隆形成能力和侵袭转移能力的变化情况。结果通过表达谱芯片结果发现Notch1在接受CGA治疗的食管癌小鼠食管组织内的表达下降,并且IHC染色结果证实这一变化。Westernblot结果显示,CGA能够抑制不同食管癌细胞中Notch1的表达,并且这种抑制作用会随着CGA处理浓度的增加而逐渐增强。采用RNAi技术敲降食管癌细胞中的Notch1后加入CGA进行克隆形成实验和Transwell实验,结果均表明,在正常表达Notch1的食管癌细胞中,经过CGA处理的细胞所形成的克隆团数目与发生侵袭转移的细胞数目远远低于对照组,而敲降Notch1之后,两组细胞所形成的克隆团数和发生侵袭转移的细胞数量未有明显的差异。结论 Notch1是CGA的作用靶点之一,CGA通过作用于Notch1进而抑制食管癌细胞的克隆形成和侵袭转移。

Objective This study aims to investigate the effect and mechanism of Notch1 in the process of chlorogenic acid(CGA)suppressing esophageal squamous cell carcinoma(ESCC).Methods Microarray assay was performed to detect the different genes in esophagus tissues of ESCC murine model treated with CGA,and immunohistochemistry(IHC)was used for verification.Different ESCC cell lines were treated with CGA,and the expression of Notch1 in these cells was detected by Western blot.RNAi was used to knock down Notch1 in ESCC cells before adding CGA treatment,then the colony formation assay and Transwell invasion assay were performed to detect the capabilities of colony formation and invasion in these cells,respectively.Results Microarray assay analysis showed that the expression of Notch1 in esophagus tissues was down-regulated in mice treated with CGA,which was confirmed by IHC staining.Inhibition of Notch1 expression in different ESCC cell lines was observed by Western blot,and the inhibitory effect was gradually enhanced with the increase of CGA concentration.Using RNAi technology to knock down Notch1 in ESCC cells and adding CGA to perform colony formation assay and Transwell invasion assay,the results showed that the number of colonies formed and cells invaded by CGA treatment was much lower than that of the control group in ESCC cells that normally express Notch1.However,in Notch1 knocked-down ESCC cells,no significant difference of the number of either colonies formed or cells invaded was observed after the treatment of CGA.Conclusion Notch1 is one target of CGA,and CGA inhibits the capabilities of colony formation and invasion of ESCC cells by acting on Notch1.