Mechanism of silencing circ-NDRG1 regulating the malignant biological behavior of oral squamous cell carcinoma cells
Objective:To investigate the expression of circ-NDRG1 in oral squamous cell carcinoma(OSCC)tissues and the effect on the malignant biological behavior of OSCC HSC-3 cells by targeting miR-1271.Methods:The TCGA database was used to analyze the expression level of circ-NDRG1 in OSCC tissues,and the correlation between its expression and patient's survival was analyzed.The expression level of circ-NDRG1 in immortalized oral keratinocyte HOK and OSCC HSC-3,SCC-25,CAL-27,Tca-8113 cells was detected by real-time fluorescence quantitative PCR(qPCR).NC inhibitor and circ-NDRG1 inhibitor were transfected into HSC-3 cells respectively by using Lipofectamine 2000(Anti-NC group and Anti-circ-NDRG1 group).The proliferation and invasion abilities of HSC-3 cells in each group were detected by MTT and Transwell chamber assay.The expression of HSC-3 cell proliferation proteins(CDK3,Cyclin C)and epithelial-mesenchymal transition(EMT)proteins(ZLF8,Notch,SIX1)was detected by Western blot.The TCGA database was used to analyze the relationship between the expression of circ-NDRG1 and miR-1271 in OSCC tissues.The dual-luciferase reporter assay was used to verify the targeting relationship between circ-NDRG1 and miR-1271.The expression of miR-1271 in HSC-3 cells in each group was detected by qPCR.Results:TCGA database analysis showed that the expression of circ-NDRG1 in OSCC tissues was significantly higher than that in adjacent tissues(P<0.01).The survival time of OSCC patients was negatively correlated with the expression level of circ-NDRG1(P<0.01).Compared with HOK cells,circ-NDRG1 was highly expressed in HSC-3,SCC-25,CAL-27,and Tca-8113 cells(P<0.01).Compared with the Anti-NC group,low expression of circ-NDRG1 could inhibit the proliferation ability(P<0.05)and invasion ability(P<0.01)of HSC-3 cells.Compared with the Anti-NC group,low expression of circ-NDRG1 could downregulate the protein expressions of CDK3,Cyclin C,ZLF8,Notch,and SIX1 in HSC-3 cells(all P<0.01).The expression level of circ-NDRG1 in OSCC tissues was negatively correlated with the expression level of miR-1271(P<0.01).circ-NDRG1 could target miR-1271(P<0.01).Compared with the Anti-NC group,low expression of circ-NDRG1 could upregulate the expression of miR-1271 in HSC-3 cells(P<0.01).Conclusion:Circ-NDRG1 is highly expressed in OSCC tissues,and silencing circ-NDRG1 can inhibit the proliferation,invasion and EMT process of OSCC cells by targeting miR-1271.
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维普
