Abstract：Objective To explore the mechanism of HOTAIR/CSTF2/CD44 axis regulating invasion and metastasis of esophageal cancer. Methods EC tissue samples and adjacent normal tissues from 43 patients with nonspecific invasive EC from September 2020 to June 2022 were collected and divided into control group and EC group. EC cell lines KYSE150, EC109 and TE-1 and human normal esophageal epithelial cell line HEEC cells were cultured and divided into control, si-HOTAIR, CSTF2 mimics and si-HOTAIR+CSTF2 mimics. Real-time fluorescence quantitative PCR was used to detect the expression of HOTAIR, CD24 and CSTF2, CCK8 was used to measure cell viability, flow cytometry was used to detect apoptosis, and then cell migration and invasion were detected. Apoptotic proteins (Ki67, PCNA, lysed caspase 3 and caspase 9) and migration proteins (VEGF and MMP-9) were detected by western blotting. Mouse models were monitored for signs of tumor growth. Results The results showed that the expression of HOTAIR and CSTF2 in EC tissues was significantly higher than that in normal tissues. Compared with HEEC cells, the expressions of HOTAIR and CSTF2 were increased in EC cell lines EC109, KYSE150 and TE-1. Compared with control, HHOTAIR knockdown significantly reduced cell proliferation, migration and invasion, and accelerated cell apoptosis. Bioinformatics analysis of CSTF2 interaction showed that there was a potential binding domain in HOTAIR transcripts, and there was a strong correlation between HOTAIR and CSTF2 expression. HOTAIR knockdown resulting in CSTF2 downregulation significantly inhibited CD24 expression, and CSTF2 elevation significantly increased CD24 expression. In vivo studies have shown that HOTAIR knockdown can reduce the expression of CSTF and CD24, thus inhibiting the growth and invasion of tumor cells. Conclusion The results showed that the expression of HOTAIR and CSTF2 in EC cells and tissues increased significantly. In vitro and in vivo studies have shown that HOTAIR knockdown promotes apoptosis and inhibits cell proliferation and invasion. HOTAIR may bind CSTF2 and indirectly regulate the expression of CD24 in EC cells.
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