Effects of circNEIL3 Targeting miR-218-5p on Proliferation, Apoptosis, Migration and Invasion of Oral Squamous Cell Carcinoma Cells
LIU Yueqiang*, XING Hongyan, FENG Jiangjie
This study aimed to investigate the effect of circNEIL3 targeting miR-218-5p on the biological behavior of oral squamous cell carcinoma cells. qRT-PCR was used to detect the expression of circNEIL3 and miR-218-5p in oral squamous cell carcinoma tissue, adjacent tissue and human oral squamous cell carcinoma cell (CAL-27, SCC-25, SCC-9, HSC-3) and normal oral keratinocyte HOK. Taking CAL-27 cells as the research object, si-circNEIL3, miR-218-5p mimics, si-NC, miR-NC were transfected into CAL-27 cells, si-circNEIL3 and anti-miR-NC, si-circNEIL3 and anti-miR-218-5p were co-transfected into CAL-27 cells. CAL-27 cell proliferation was detected by MTT assay; CAL-27 cell apoptosis was detected by flow cytometry; the invasion and migration of CAL-27 cells were detected by Transwell. Dual-luciferase reporter assay was used to detect the targeting relationship between circNEIL3 and miR-218-5p. Western blot was used to detect the expression of Bax, Bcl-2, caspase-3, and cleaved-caspase-3 protein. Compared with adjacent tissues, the expression of circNEIL3 in oral squamous cell carcinoma tissues was increased (P<0.01), while the expression of miR-218-5p was decreased (P<0.01); compared with HOK cells, the expression of circNEIL3 in human oral squamous cell carcinoma cells (CAL-27, SCC-25, SCC-9, HSC-3) was increased (P<0.05), and the expression of miR-218-5p was decreased (P<0.05). Compared with transfection of si-NC or miR-NC, transfection of si-circNEIL3 or miR-218-5p mimics increased cell proliferation inhibition rate, apoptosis rate, and Bax, caspase-3, and cleaved-caspase-3 protein levels (P<0.01), decreased Bcl-2 protein level (P<0.01), and reduced the number of migrating and invasive cells (P<0.01). circNEIL3 could negatively regulate the expression of miR-218-5p. Compared with co-transfection of si-circNEIL3 and anti-miRNC, after co-transfection with si-circNEIL3 and anti-miR-218-5p, the cell proliferation inhibition rate, apoptosis rate and the protein levels of Bax, caspase-3, and cleaved-caspase-3 were reduced (P<0.01), while the number of migrating and invasive cells was increased (P<0.01), and the protein level of Bcl-2 was increased (P<0.01). circNEIL3 can inhibit the proliferation, migration and invasion of oral squamous cell carcinoma cells and promote cell apoptosis by targeting and negatively regulating miR-218-5p.
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