Effect of Silencing circCCDC66 on the Proliferation and Apoptosis of Esophageal Cancer Cells Eca-109 by Targeting miR-129-5p
LI Dongming*, XIANG Kemin, WEI Yunjia
In this paper, the effect of silencing circCCDC66 on the proliferation and apoptosis of Eca-109 in esophageal cancer cells and its possible mechanism were discussed. The expression of circCCDC66, miR-129- 5p and HMGB1 in esophageal cancer tissues and adjacent tissues were detected by qRT-PCR and Western blot. Human esophageal cancer cells Eca-109 were cultures in vitro, si-NC, si-circCCDC66, miR-NC, miR-129-5p mimic, si-circCCDC66+miR-129-5p inhibitor were transfected into Eca-109 cells, respectively. The expression of circCCDC66, miR-129-5p and HMGB1 in cells were detected by qRT-PCR and Western blot, respectively. Cell proliferation, colony formation and apoptosis rate were detected by CCK-8 assay, plate clone formation as-say and flow cytometry, respectively. The targeting relationship between circCCDC66 and miR-129-5p, and the targeting relationship between miR-129-5p and HMGB1 were verified by dual luciferase report experiment. The expression levels of circCCDC66 and HMGB1 in esophageal cancer tissues were higher than those in paracancer tissues (P<0.05), while the expression of miR-129-5p was lower than those in paracancer tissues (P<0.05). After transfection with si-circCCDC66 or miR-129-5p mimic, the expression of miR-129-5p, the rate of apoptosis, and the inhibition rate of cell proliferation were increased (P<0.05), while the protein level of HMGB1 and the number of colonies formed were decreased (P<0.05). circCCDC66 could target and regulate the expression of miR129-5p, and HMGB1 is the target gene of miR-129-5p. Co-transfection of si-circCCDC66 and miR-129-5p inhibitor could reduce the effect of transfection of si-circCCDC66 on the proliferation, clone formation and apoptosis of Eca-109 cells. Silencing circCCDC66 can reduce the proliferation and clonal formation ability of esophageal cancer cells and induce cell apoptosis by targeting the expression of miR-129-5p/HMGB1.
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