24 March 2020 : Laboratory Research
PIK3C3 Acts as a Tumor Suppressor in Esophageal Squamous Cell Carcinoma and Was Regulated by MiR-340-5p
Xiaobo Wang1ABCEFG, Min Gu1BCD, Yongjian Ju1DEFG, Juying Zhou2BCDFG*DOI: 10.12659/MSM.920642
Med Sci Monit 2020; 26:e920642
Abstract
BACKGROUND: Esophageal squamous cell carcinoma (ESCC), a major histological subtype of esophageal cancer, is a common cause of tumor-related deaths in the world. Due to the lack of understanding of the pathogenesis of ESCC, its clinical treatment is still a big challenge. In the present study, we aimed to identify an ESCC-related gene in the GEO dataset, and to explore its function and mechanism in ESCC.
MATERIAL AND METHODS: The GSE dataset (GSE100492) consisting of 10 samples was analyzed using GEO2R for identifying the differentially expressed genes between ESCC and normal samples. Expression levels of mRNA and miRNA in ESCC tissues and cells were detected via quantitative real-time polymerase chain reaction. Protein expression was analyzed by western blot. Cell proliferation viability was determined through MTT and colony formation. Cell distribution and apoptosis was detected by flow cytometry. MiRNA target prediction was analyzed by bioinformatics. The interplay between miR-340-5p and PIK3C3 was validated by dual-luciferase reporter assay.
RESULTS: PIK3C3 was lowly expressed in ESCC tissue and indicated a poor prognosis in patents. Overexpression of PIK3C3 in vitro repressed cell proliferation of KYSE-150 and TE-12 cells. Moreover, PIK3C3 overexpression was demonstrated to enhance the sensitivity of KYSE-150 and TE-12 cells to irradiation. In addition, miR-340-5p was revealed to directly bind and negatively modulate PIK3C3 expression in ESCC. Blockage of miR-340-5p promoted ESCC cell proliferation, while rescue of PIK3C3 reversed this effect. MiR-340-5p was highly expressed in ESCC tissue and it exhibited a negative correlation with PIK3C3 expression.
CONCLUSIONS: MiR-340-5p functioned as an oncogene of ESCC by directly binding and repressing the expression of PIK3C3.
Keywords: Cell Proliferation, Esophageal Neoplasms, MicroRNAs, Base Sequence, Class III Phosphatidylinositol 3-Kinases, Down-Regulation, esophageal squamous cell carcinoma, Gene Expression Regulation, Neoplastic, Genes, Tumor Suppressor, Middle Aged, Radiation, Ionizing
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