Objective This study aims to investigate the effect and mechanism of Notch1 in the process of CHA 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 CHA, and immunohistochemistry (IHC) was used for verification. Different ESCC cell lines were treated with CHA, 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 CHA 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 CHA, 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 CHA concentration. Using RNAi technology to knock down Notch1 in ESCC cells and adding CHA to perform colony formation assay and Transwell invasion assay, the results showed that the number of colonies formed and cells invaded by CHA 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 CHA.

Conclusion Notch1 is one target of CHA, and CHA inhibits the capabilities of colony formation and invasion of ESCC cells by acting on Notch1.

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