Objective To investigate the biological roles and potential regulatory mechanisms of microRNA-4465 (miR-4465) in colon cancer cells.
Methods qPCR was used to detect the miR-4465 levels in SW480, HT-29 and HCT-116 cells; then miR-4465 was overexpressed in HCT-116 cells, and cell viability and apoptosis were detected by MTT assay and flow cytometry, respectively; the Caspase-3 activity was measured by a kit; the cell invasion was detected by Transwell assay; the targeting relation between miR-4465 and HMGA1, HMGA2, EZH2 were determined by the luciferase reporter gene assay. The protein and mRNA expression levels of HMGA1, HMGA2 and EZH2 were detected by Western blot and qPCR, respectively.
Results The expression of miR-4465 was significantly down-regulated in SW480, HT-29 and HCT-116 cells (P < 0.05). Overexpression of miR-4465 reduced the viability of HCT-116 cells, increased cell apoptosis, enhanced Caspase-3 activity and suppressed cell invasion (all P < 0.05). MiR-4465 directly targeted 3'-UTR of HMGA1, HMGA2 or EZH2, thereby regulating their expression negatively.
Conclusion MiR-4465 overexpression could reduce the viability of colon cancer cells, promote cell apoptosis and suppress cell invasion, which may be related to the negative regulation of HMGA1, HMGA2 or EZH2.