Abstract:
Objective: To investigate the effect of SerpinA5 on the proliferation, apoptosis, migration and invasion of esophageal squamous cell carcinoma (ESCC) cells and its possible molecular mechanism. Methods: The expression level of SerpinA5 gene between various tumors and adjacent normal tissues was analyzed by TIMER2.0 database. Western blot was used to detect the expression of SerpinA5 in two human ESCC cell lines (KYSE150 and TE-1) and esophageal epithelial cell line Het-1A. The SerpinA5 overexpression KYSE150 cell line was constructed by lentivirus, and the overexpression efficiency was detected by Western blot. The effects of SerpinA5 overexpression on the proliferation, apoptosis, migration and invasion of ESCC cells were detected by CCK8, plate cloning assay, flow cytometry, wound healing assay and Transwell invasion chamber. The SerpinA5 overexpression cell line was subcutaneously injected into the right axilla of nude mice to establish a subcutaneous xenograft model. The tumor growth was observed, and the volume and mass of the tumor were measured. The cell proliferation level of subcutaneous xenograft tumor in nude mice was detected by IHC. Co-immunoprecipitation (Co-IP) and Western Blot were used to determine the interaction between SerpinA5 and Fn. Western blot was used to detect the expression of Fn/Integrin-β1 signaling pathway-related proteins (Fn, Integrin-β1, FAK and p-FAK) in transplanted tumors. Results: SerpinA5 was lowly expressed in ESCC tissues and cell lines. Overexpression of SerpinA5 in ESCC cells can significantly inhibit cell proliferation, migration and invasion, and promote cell apoptosis. In the subcutaneous xenograft experiment in nude mice, the tumor volume and weight of SerpinA5 overexpression group were smaller than those of negative control group. IHC results showed that overexpression of SerpinA5 significantly inhibited the proliferation of ESCC cells in tumor tissues. Co-IP confirmed the interaction between SerpinA5 and Fn. At the same time, Western blot results showed that the expression levels of Fn, Integrin-β1 and p-FAK in the Fn/Integrin-β1 signaling pathway of ESCC cells in the subcutaneous xenograft of nude mice were significantly decreased after overexpression of SerpinA5. Conclusions: Serpin A5 may inhibit the proliferation, migration, invasion and promote apoptosis of ESCC cells by regulating Fn/Integrin-β1 signaling pathway.
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