Molecular mechanism of circBACH1 regulating the proliferation, migration and invasion of oral squamous cell carcinoma HSC3 cells by targeting miR-4500

ObjectiveTo investigate the effect of circBACH1 on the biological behavior of oral squamous cell carcinoma HSC3 cells and its possible mechanism.

MethodsHSC3 cells were cultured in vitro and divided into si-NC group, si-circBACH1 group, miR-NC group, miR-4500 group, si-circBACH1+anti-miR-NC group, si-circBACH1+anti-miR-4500 group, which was transfected into HSC3 cells with si-NC, si-circBACH1, miR-NC, miR-4500 mimics, si-circBACH1 plus anti-miR-NC, si-circBACH1 plus anti-miR-4500, respectively.qRT-PCR was used to detect the expression of circBACH1 and miR-4500 in oral squamous cell carcinoma tissue, its adjacent tissue and HSC3 cells in the groups.CCK-8 and plate clone formation assay were applied to determine cell proliferation and clone formation ability.Scratch assay and Transwell chamber assay were employed to analyze the cell migration and invasion ability.The dual luciferase reporter gene experiment was performed to evaluate the targeted regulation effect of circBACH1 on miR-4500.Western blotting was used to detect the protein expression of E-cadherin and N-cadherin.

ResultsCompared with the adjacent tissue, the expression of circBACH1 in oral squamous cell carcinoma tissue was significantly increased (P < 0.01), and the expression of miR-4500 was significantly decreased (P < 0.01).Compared with the si-NC group, the viability, number of cell clones and invaded cells, wound healing rate and protein level of N-cadherin of HSC3 cells in the si-circBACH1 group were significantly reduced (P < 0.01), and the protein level of E-cadherin was significantly increased (P < 0.01).Compared with the miR-NC group, the viability, number of cell clones and invaded cells, wound healing rate and protein level of N-cadherin of HSC3 cells in the miR-4500 group were significantly decreased (P < 0.01), and the protein level of E-cadherin was significantly increased (P < 0.01).The luciferase activity of HSC3 cells in the WT-circBACH1 plus miR-4500 mimics co-transfection group was significantly lower than that in the WT-circBACH1 plus miR-NC co-transfection group (P < 0.01).The results of qRT-PCR showed that compared with the pcDNA group, the expression of miR-4500 in the pcDNA-circBACH1 group decreased (P < 0.05);compared with si-NC group, the expression of miR-4500 in si-circBACH1 group increased (P < 0.05).Compared with the si-circBACH1+anti-miR-NC group, the viability, number of cell clones and invaded cells, wound healing rate and protein level of N-cadherin of HSC3 cells in the si-circBACH1+anti-miR-4500 group were significantly increased (P < 0.01), and the protein level of E-cadherin was significantly decreased (P < 0.01).

ConclusionsInterfering with the expression of circBACH1 can inhibit the proliferation, clone formation, migration and invasion of oral squamous cell carcinoma HSC3 cells by negatively regulating the expression of miR-4500.