MiR-20b-5p functions as tumor suppressor microRNA by targeting cyclinD1 in colon cancer

Abstract

ABSTRACT MicroRNA functions as an oncogenic regulator or tumor suppressor in various human tumors. Although bioinformatics analysis suggested that miRNA-20b-5p may be associated with the tumorigenesis, its role in colon cancer remains elusive. To investigate the role of miRNA-20b-5p, HCT116 cell, a human colon cancer cell line used in therapeutic research and drug screenings, was chosen as a model system for our in vitro studies. We first carried out bioinformatics and microarray analysis. To gain further mechanism insight, flow cytometry was performed to determine cell apoptosis and cell cycle, and western blot or immunohistochemistry were employed to check the expression of CCND1/CDK/FOXM1 axis in HCT116 cells. In addition, wound-healing migration assay and transwell assay were conducted to uncover the effect of miR-20b-5p on tumor migration and invasion. Finally, we examined the role of miR-20b-5p by subcutaneous xenograft mouse models. Our data have shown that miRNA-20b-5p inhibited the cell cycle, migration, and invasion in HCT116 cells, but had no effect on cell apoptosis. CyclinD1 (CCND1) was identified as a direct target of miR-20b-5p. Overexpression of miRNA-20b-5p downregulated CCND1 level in HCT-116 cells. Mechanically, the inhibition of cell cycle, migration, and invasion of CC cells mediated by miRNA-20b-5p are through regulating the CCND1/CDK4/FOXM1 axis. Furthermore, miRNA-20b-5p inhibited the tumorigenesis in Balb/c nude mice CC xenograft models. Our data demonstrated that miR-20b-5p may serve as a tumor suppressor in colon cancer by negatively regulating CCND1, implying that miR-20b-5p could be a potential therapeutic target for the treatment of colon cancer.