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Archive > Volume 30 Issue 2 > 2023,30(2):114-122. DOI:10.3872/j.issn.1007-385X.2023.02.003 Prev Next
Basic Research
hsa_circ_0140180 affects the migration and invasion of esophageal squamous cell carcinoma cells by regulating miR-1287-5p
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Abstract:
Objective: To investigate the expression level of hsa_circ_0140180 in esophageal squamous cell carcinoma (ESCC) cells,as well as its effect on malignant biological behavior of ESCC cells and the possible molecular mechanisms. Methods: Six pairs of ESCC tissues and corresponding paracarcinoma tissues resected at the Department of Thoracic Surgery, Central Hospital of Nanchong City between November 2018 and March 2019 were collected for whole-transcriptome sequencing, and hsa_circ_0140180 at a low expression in ESCC tissues was screened out. The TE-1 and KYSE30 cell lines overexpressing hsa_circ_0140180 were established.qPCR was used to detect the expression of hsa_circ_0140180 in human normal esophageal epithelial cells and ESCC cells, as well as the expression of miR-1287-5p in TE-1 and KYSE30 cells overexpressing hsa_circ_0140180. CCK-8 and FCM were used to detect theeffect of hsa_circ_0140180 overexpression on the proliferation and cell cycle of TE-1 and KYSE30 cells. Scratch assay and Transwellassay were used to assess the effects of hsa_circ_0140180 overexpression on migration and invasion of TE-1 and KYSE30 cells. Dual-luciferase reporter assay was used to confirm the targeting relationship between hsa_circ_0140180 and miR-1287-5p. WB assay was used to detect the expression of EMT-related proteins and the phosphorylation level of the PI3K-Akt pathway in TE-1 and KYSE30 cells after hsa_circ_0140180 overexpression. Results: Transcriptome sequencing and qPCR results showed that hsa_circ_0140180 was lowly expressed in ESCC tissues and cells (P<0.05 or P<0.01) and confirmed its closed-loop molecular signature and localization in the cytoplasm. Overexpression of hsa_circ_0140180 could significantly inhibit the migration and invasion of ESCC cells (all P<0.05) but had no significant effect on the proliferation and cell cycle. The results of dual-luciferase reporter gene assay demonstrated that hsa_circ_0140180 targets miR-1287-5p and negatively regulates its expression (P<0.001). Overexpression of hsa_circ_0140180 could significantly increase the expression of E-cadherin and decrease the expression of Snail (both P<0.05) and the phosphorylation level of the PI3K-Akt pathway (P<0.01, P<0.001) in TE-1 and KYSE30 cells. Conclusion: hsa_circ_0140180 is lowly expressed in ESCC cells and tissues; it reduces the phosphorylation levels of PI3K-Akt pathway and suppresses EMT process possibly through regulation of miR-1287-5p expression, thus affecting the migration and invasion abilities of ESCC cells.
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