Inhibition of microRNA-383 promotes apoptosis of human colon cancer cells by upregulation of caspase-2 gene expression

Purpose : To investigate microRNA-383 (miR-383) as a therapeutic target for the management of colon cancer. Methods : Total RNA was isolated using RNeasy RNA isolation kit according to the manufacturer’s instructions. cDNA was synthesized using RevertAid cDNA synthesis kit. Expression analysis was carried out by quantitative real-time polymerase chain reaction (RT-PCR). Cell proliferation was examined using CellTiter 96 AQueous One Solution Cell Proliferation Assay system, while apoptosis was detected by 4',6-diamidino-2-phenylindole (DAPI) and annexin V/PI double staining followed by flow cytometry. The miR-383 target was delimited using TargetScan software. Protein expression analysis was carried out by western blotting. Results : The results indicate that miR-383 was highly expressed in colon cancer cells. Down-regulation of miR-383 inhibited cancer cell proliferation, and promoted apoptosis and cell cycle arrest. Furthermore, in silico analysis revealed caspase-2 gene to be the downstream target of miR-383, a finding that was further confirmed by western blotting. Conclusion : The results reveal that miR-383 may be an important target to tackle the increasing incidence of colon cancer. Thus, drugs that target miR-383 and inhibit its expression can potentially be developed for the treatment of colon cancer. Keywords : MicroRNA, Colon cancer, Cell proliferation, Apoptosis, Protein expression