Volume 29, Issue 3
DOI: 10.24205/03276716.2020.746
Effects of circular ribonucleic acid G-protein subunit beta 1 on migration, invasion and apoptosis of colon cancer cells
Abstract
Objective: To explore the effects of circular ribonucleic acid G-protein subunit beta 1 (circGNB1) on the migration, invasion and apoptosis of colon cancer cells and its molecular mechanism.
Methods: The small-interfering RNA-negative control (si-NC) group, si-circGNB1 group, pcDNA-NC group, pcDNA-circGNB1 group, miR-NC group, miR-432-5p group, si-circGNB1 anti-miR-NC group, and si-circGNB1 anti-miR-432-5p group were set up. The expression levels of circGNB1 and miR-432-5p were determined via real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the protein expressions of matrix metalloproteinase-2 (MMP2), MMP9 and cleaved-caspase-3 were determined via Western blotting. The cell migration and invasion were detected using Transwell assay, and the apoptosis was detected using flow cytometry. The targeting relationship between circGNB1 and miR-432-5p was verified through dual-luciferase reporter assay.
Results: Compared with normal human colonic epithelial cells FHC, colon cancer cell lines HCT116, HT29 and SW480 had a significantly increased expression of circGNB1 and a significantly decreased expression of miR-432-5p (P<0.05). After low expression of circGNB1 or high expression of miR-432-5p, the expression levels of MMP2 and MMP9 significantly declined, the expression level of cleaved-caspase-3 significantly rose, the number of migrating and invading cells was significantly decreased, and the apoptosis rate was significantly increased (P<0.05). CircGNB1 regulated miR-432-5p in a targeted manner, and lowly-expressed miR-432-5p partially reversed the effects of lowly-expressed circGNB1 on the migration, invasion and apoptosis of HCT116 cells.
Conclusion: Lowly-expressed circGNB1 may inhibit the migration and invasion and promote the apoptosis of HCT116 cells through up-regulating miR-432-5p.
Keywords
CircGNB1, miR-432-5p, colon cancer, migration, invasion, apoptosis.