hsa_circ_0011946 regulates the proliferation,migration and invasion of cervical cancer cells via targeting miR-767-3p
Objective To investigate the role of hsa_circ_0011946 targeting miR-767-3p in the progression of cervical cancer.Methods The expression levels of hsa_circ_0011946 and miR-767-3p in cervical cancer tissues,adjacent tissues,immortalized human cervical epithelial cells(H8)and cervical cancer cells(SiHa,HeLa,and Cashi)were determined by RT-qPCR.SiHa cells were transfected with si-NC,si-hsa_circ_0011946,miR-NC,miR-767-3p,pcDNA,pcDNA-hsa_circ_0011946,anti-miR-NC+si-hsa_circ_0011946,anti-miR-767-3p+ si-hsa_circ_0011946,respectively,as the si-NC group,the si-hsa_circ_0011946 group,the miR-NC group,the miR-767-3p group,the pcDNA group,the pcDNA-hsa_circ_0011946 group,the anti-miR-NC+si-hsa_circ_0011946 group,and the anti-miR-767-3p+si-hsa_circ_0011946 group,and the untransfected cells were used as the NC group.The expression of hsa_circ_0011946 and miR-767-3p of SiHa cells in each group was detected by RT-qPCR,the cell viability was detected by CCK-8,the number of cell clone was detected by plate clone formation assay,the cell migration and invasion numbers were detected by Transwell assay,and the protein expression of MMP-2 and MMP-9 was detected by Western blot.The binding site of miR-767-3p to hsa_circ_0011946 was predicted by circular RNA interactome,and the targeted relationship between hsa_circ_0011946 and miR-767-3p was analyzed by dual luciferase reporter assay.Results Compared with the adjacent tissues,the expression level of hsa_circ_0011946 in the cervical cancer tissues was significantly increased(P<0.05),while the expression level of miR-767-3p was significantly decreased(P<0.05).Compared with H8 cells,the expression levels of hsa_circ_0011946 in SiHa,HeLa and Caski cells were significantly increased(P<0.05),while the expression levels of miR-767-3p were signifi-cantly decreased(P<0.05).Compared with the NC group,the expression level of hsa_circ_0011946,absorbance(A)value,number of clone,number of migration,number of invasion and the protein expression levels of MMP-2 and MMP-9 were significantly decreased in SiHa cells in the miR-767-3p group(P<0.05).Compared with the miR-NC group,the expression level of miR-767-3p in SiHa cells in the miR-767-3p group was significantly increased(P<0.05),and the cell A value,number of clone,number of migration,number of invasion and the protein expression levels of MMP-2 and MMP-9 were significantly decreased(P<0.05).Compared with the pcDNA group,the expression level of miR-767-3p in SiHa cells in the pcDNA-hsa_circ_0011946 group was significantly decreased(P<0.05),and the expression level of hsa_circ_0011946 was significantly increased(P<0.05).The relative luciferase activity of SiHa cells co-transfected with miR-767-3p mimics and WT-hsa_circ_0011946 was lower than that of co-transfected with miR-NC and WT-hsa_circ_0011946(P<0.05).hsa_circ_0011946 bound to miR-767-3p directly and specifically.Compared with the anti-miR-NC+si-hsa_circ_0011946 group,the expression level of miR-767-3p in SiHa cells in the anti-miR-767-3p+si-hsa_circ_0011946 group was significantly decreased(P<0.05),and the cell A value,number of clone,number of migration,number of invasion and protein expression levels of MMP-2 and MMP-9 were significantly increased(P<0.05).Conclusion Interfering hsa_circ_0011946 can inhibit the proliferation,migration and invasion of cervical cancer cells through targeted up-regulation of miR-767-3p.