CircCDK1 knockdown reduces CDK1 expression by targeting miR-489-3p to suppress the development of breast cancer and strengthen the sensitivity of Tamoxifen.

Abstract

Circular RNAs (circRNAs) are implicated with the progression of multiple cancers, including breast cancer. Besides, circRNA dysregulation is involved in the chemoresistance of cancer development. This study aimed to investigate the role of circRNA-cyclin dependent kinase 1 (circCDK1) in breast cancer. Quantitative real-time PCR (qPCR) and western blot were applied for expression analysis. Cell viability was determined by the cell counting kit-8 (CCK-8). Cell proliferation was evaluated by CCK-8, colony formation and 5-ethynyl-2'-deoxyuridine assays. Cell apoptosis was assessed by flow cytometry and the activities of caspase3 and caspase9. The potential binding between miR-489-3p and circCDK1 or CDK1 was verified by RNA immunoprecipitation assay, dual-luciferase reporter assay and pull-down assay. Animal models were constructed to explore the role of circCDK1 in vivo. CircCDK1 was overexpressed in Tamoxifen-resistant breast cancer cells, LCC2 and LCC9. The expression of circCDK1 in tumor tissues with Tamoxifen resistance was higher than that in tissues without Tamoxifen resistance. CircCDK1 knockdown strengthened the sensitivity of Tamoxifen in LCC2 and LCC9 cells and reduced Tamoxifen IC50. The downregulation of circCDK1 inhibited LCC2 andLCC9 cell proliferation and promoted cell apoptosis. CDK1 was the parent gene of circCDK1 and circCDK1 positively regulated CDK1 expression by targeting miR- 489-3p. CDK1 overexpression reversed the effects of circCDK1 knockdown. MiR-489-3p inhibition also reversed the effects of circCDK1 knockdown. CircCDK1 knockdown was verified to enhance Tamoxifen sensitivity in animal models. CircCDK1 knockdown enhanced the sensitivity of Tamoxifen in breast cancer cells and suppressed cell growth and survival by depleting CDK1 expression via releasing miR- 489-3p.