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Research Paper Volume 12, Issue 14 pp 14157—14173
Circular RNA hsa_circ_0000073 contributes to osteosarcoma cell proliferation, migration, invasion and methotrexate resistance by sponging miR-145-5p and miR-151-3p and upregulating NRAS
Figure 5. The regulatory relationship among hsa_circ_0000073, miR-145-5p, miR-151-3p and NRAS in OS. (A) The binding sites between hsa_circ_0000073 and miR-145-5p or miR-151-3p. (B) qRT-PCR analyses of miR-145-5p and miR-151-3p in OS and paired paratumor tissues. (C) MiR-145-5p and miR-151-3p expression in hFOB 1.19 and OS cells was monitored by qRT-PCR assay. (D) The correlativity between hsa_circ_0000073 and miR-145-5p or miR-151-3p expression in OS tissues. (E) qRT-PCR analyses of miR-145-5p and miR-151-3p in hsa_circ_0000073-overexpressing and hsa_circ_0000073-silenced MG-63 and U2OS cells. (F–G) MG-63 and U2OS cells were cotransfected with the hsa_circ_0000073 plasmid and miR-145-5p or miR-151-3p mimics, and the levels of hsa_circ_0000073 and NRAS were determined through qRT-PCR. (H, I) Western blot analysis of NRAS expression in MG-63 and U2OS cells after transfection with miR-145-5p or miR-151-3p mimics and inhibitors. (J) The luciferase intensities between NRAS and miR-145-5p or miR-151-3p were assessed via dual-luciferase reporter gene assay. (K–M) qRT-PCR and western blot assays were performed to verify the impacts of hsa_circ_0000073 and miR-145-5p or miR-151-3p on NRAS expression. *P<0.05.