circ_0015756 Affects Cell Proliferation, Apoptosis and Migration by Regulating miR-515-5p/HMGB3 Axis in Lung Cancer
XU Jun1, ZHU Chengying1, WANG Dejun2*
In order to investigate the effect of circ_0015756 (circular RNA 0015756) on the proliferation, apoptosis and migration of lung cancer cells and its underlying mechanism, RT-qPCR was applied to analyze the expression of circ_0015756 and microRNA (miR)-515-5p in lung cancer tissues and adjacent tissues. Meanwhile, si-circ_0015756 (circ_0015756 small interfering RNA), miR-515-5p mimic, si-circ_0015756+miR-515-5p inhibitor were transfected into lung cancer cell A549, respectively. The proliferation ability of A549 was assessed by methyl thiazolyl tetrazolium method and plate cloning assay. The apoptosis rate of A549 was measured by flow cytometry, and the migration ability of A549 was evaluated by scratch healing and Transwell test. Western blot was used to assess the expression level of HMGB3 (high mobility group protein 3). The targeting relationships between circ_0015756 and miR-515-5p, miR-515-5p and HMGB3 were verified by dual luciferase assays. The results showed that the relative level of circ_0015756 in lung cancer tissue was significantly higher than that in adjacent tissues (P<0.05), while the relative level of miR-515-5p was significantly lower than that in adjacent tissues (P<0.05). After interfering with the expression of circ_0015756, the cell proliferation inhibition rate, apoptosis rate, and miR-515-5p relative levels of A549 cells were significantly increased (P<0.05), and the colony forming numbers, migration distance, migration cell numbers and relative levels of HMGB3 protein were significantly reduced (P<0.05). After overexpression of miR-515-5p, the proliferation inhibition rate and apoptosis rate of A549 cells were significantly increased (P<0.05). The number of colonies formed, migration distance, migration cell numbers and the relative level of HMGB3 protein were significantly reduced (P<0.05). miR-515-5p inhibition significantly reduced the effect of interfering with circ_0015756 on the proliferation, colony formation, migration and HMGB3 protein expression of A549 cells (P<0.05). circ_0015756 directly bound to miR-515-5p, and miR-515-5p directly bound to HMGB3. In conclusion, interference circ_0015756 inhibited the proliferation and migration, and induced cell apoptosis of lung cancer cells by targeting and up-regulating miR-515-5p/HMGB3 axis.
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