Abstract:

Objective To investigate the effect of Notch1^P1641S mutation on oral squamous cell carcinoma. Methods CAL27 and HSC3 were cultured in vitro. Notch1^N1 (N1-N1), Notch1^WT (N1-WT) and Notch1^P1641S (N1-P1641S) locus mutant plasmids were constructed. Plasmids were transfected into cell lines and the transfection efficiency was examined by qPCR and Westen blot assay. The effects of Notch1 mutation on proliferation, invasion and migration of CAL27 and HSC3 cells were investigated by CCK-8 assay, plate colony formation assay, Scratch assay, EdU assay and Transwell invasion assay. Western blot was used to detect the expression of cellular pathways. The effect of mutant plasmids on oral squamous carcinoma was further verified by subcutaneous tumorigenesis assay in nude mice. Results The results of the phenotypic assay showed that the proliferation, migration and invasion ability of oral squamous carcinoma cell lines in the N1-P1641S group were enhanced after transfection with the mutant plasmid compared to the N1-N1 and N1-WT groups. The results of Westen blot experiments showed that the expression of PI3K/Akt-related indicators was upregulated in the N1-P1641S group after transfection with the mutant plasmid. Subcutaneous tumorigenesis in nude mice showed an increase in tumor size after transfection with the mutant plasmid. The expression of Ki67, a proliferation-related indicator, was enhanced. Conclusion Notch1^P1641S expression site mutation promotes proliferation of oral squamous carcinoma cells through PI3K/Akt signaling pathway.

Key words: oral squamous cell carcinoma, Notch1, mutation, PI3K