Circular RNA circANKIB1 promotes the progression of osteosarcoma by regulating miR-217/PAX3 axis

Circular RNAs (circRNAs) have been discovered to exert essential roles in human cancers, including osteosarcoma (OS). The aim of this study was to investigate the exact roles and regulatory mechanism of circRNA ankyrin repeat and IBR domain containing 1 (circANKIB1) in OS.

Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression levels of circANKIB1, microRNA-217 (miR-217) and paired box 3 (PAX3). Cell proliferation was assessed by colony formation assay. Cell cycle distribution and apoptosis rate were determined by flow cytometry analysis. Wound healing assay and transwell assay were employed to evaluate cell migration and invasion abilities. Western blot assay was used to analyze the protein levels of PAX3, E-cadherin and Vimentin. Targeting relationship between miR-217 and circANKIB1 or PAX3 was predicted by Circular RNA Interactome or TargetScan and demonstrated by dual-luciferase reporter assay. The mice xenograft model was established to confirm the role of circANKIB1 in vivo.

CircANKIB1 and PAX3 were high-expressed, whereas miR-217 was low-expressed in OS tissues and cells. Knockdown of circANKIB1 inhibited the progression of OS by reducing cell proliferation, migration, invasion, and tumor growth (in vivo), and inducing apoptosis. MiR-217 was a direct target of circANKIB1, and its inhibition reversed the inhibitory effect of circANKIB1 knockdown on the progression of OS cells. Moreover, PAX3 was a direct target of miR-217, and miR-217 exerted the anti-tumor role in OS cells by targeting PAX3. Furthermore, circANKIB1 positively regulated PAX3 expression by sponging miR-217.

Knockdown of circANKIB1 suppressed OS progression by upregulating miR-217 and downregulating PAX3, which might provide a novel insight into the pathogenesis of OS.