Mechanism of hsa_circ_0013058 in promoting invasion and migration of esophageal squamous cell carcinoma
Objective To investigate the role of hsa_circ_0013058 in esophageal squamous cell carcinoma(ESCC)and the mechanism in promoting the invasion and migration of ESCC cells.Meth-ods ESCC tissues and adjacent normal tissues of 75 patients were collected.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and RNA in situ hybridization(RISH)were used to detect the expression of hsa_circ_0013058,and the relationship between hsa_circ_0013058 and clinicopathological data of ESCC patients was analyzed.The invasion and migration ability of ESCC cells were detected by scratch test and Transwell test.The target gene of hsa_circ 0013058 was predicted by bioinformatics.The regulation of hsa_circ 0013058 on downstream genes and its effect on invasion and migration of ESCC cells were detected by Rescue experiment.Results The results of qRT-PCR showed that the expression level of hsa_circ_0013058 in the ESCC tissues was significantly higher than that in the adjacent normal tissues(t=5.078,P＜0.05).The expression level of hsa_circ_0013058 in KYSE70 was significantly higher than that in human normal esophageal epithelial cells HTT-1A(P＜0.05).At the RNA level,hsa_circ_0013058 was negatively correlated with miR-548p(r=-0.254 2,P＜0.05).The positive expression rate of hsa_circ_0013058 RNA in the ESCC tissues was 72.00％,which was significantly higher than 17.33％in the adjacent normal tis-sues(x2=6.862,P＜0.05).The positive expression of hsa_circ_0013058 was closely related to the depth of tumor invasion,differentiation degree and lymph node metastasis(P＜0.05).Scratch test results showed that overexpression of hsa_circ_0013058 enhanced the migration ability of KYSE70 cells(P＜0.05);knockdown of hsa_circ_0013058 expression decreased the migration a-bility of KYSE70 cells(P＜0.05).After overexpression of hsa_circ_0013058,the number of inva-sion and migration cells of ESCC increased,and after knockdown of hsa_circ_0013058 expression,the number of invasion and migration cells decreased(P＜0.05).The qRT-PCR confirmed that hsa_circ 0013058 could regulate the expression of microRNA-548p(miR-548p).Dual luciferase report-er gene assay showed that miR-548p was specifically bound to LPAR3,which was the direct target gene of miR-548p.Rescue experiment results indicated that hsa_circ_0013058 regulated the inva-sion and migration of ESCC cells through the miR-548p/LPAR3 signaling axis.Conclusion In ESCC,hsa_circ 0013058 inhibits the expression of miR-548p,thereby activating the LPAR3 signa-ling pathway and promoting the invasion and migration of ESCC cells.
万方数据
维普
