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.

Methods 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 tissues (χ²=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 ability of KYSE70 cells (P < 0.05). After overexpression of hsa_circ_0013058, the number of invasion 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 reporter 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 invasion 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 signaling pathway and promoting the invasion and migration of ESCC cells.