CircDCAF6 regulates the miR-196a/IGF2BP3 axis to promote proliferation and inhibit apoptosis in bovine myoblasts.

Abstract

This study investigates the role of circDCAF6 in regulating the proliferation and apoptosis of bovine myoblasts, focusing on its interaction with bta-miR-196a and IGF2BP3. Using online prediction tools like TargetScan and miRanda, we identified circDCAF6 as a target for bta-miR-196a, bta-miR-196b, and bta-miR-219-3p. Experimental results showed that interference with circDCAF6 significantly increased the expression of bta-miR-196a and miR-196b, while miR-219-3p levels remained unchanged. Following these findings, we confirmed the direct targeting relationship between circDCAF6 and bta-miR-196a using a dual luciferase reporter system and RNA pull-down experiments. Subsequent analysis revealed that circDCAF6 co-transmutation with bta-miR-196a countered the inhibitory effect of the bta-miR-196a mimic on cell proliferation marker genes (CCNA1, CCNA2, MCM6) and restored S-phase cell proportions. Additionally, circDCAF6 diminished the pro-apoptotic effects of bta-miR-196a by reducing apoptosis marker gene expression (Caspase3, Caspase6) and the proportion of early apoptotic cells. We also identified IGF2BP3 as a target of bta-miR-196a, with verification through dual luciferase assays, RT-qPCR, and Western blots. Further research indicated that interfering with IGF2BP3 significantly reduced cell proliferation and increased apoptosis, characterized by lower expression of proliferation markers and higher levels of apoptosis markers. Co-transfer experiments of siRNA for circDCAF6 and IGF2BP3 showed that circDCAF6 could mitigate the inhibitory effects caused by IGF2BP3 interference. In summary, this study highlights the critical role of circDCAF6 in bovine myoblast proliferation and apoptosis via the bta-miR-196a/IGF2BP3 axis, offering insight into muscle development and disease mechanisms.