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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

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Figure 1. Hsa_circ_0000073 was identified in a screen and was highly expressed in OS. (A) Hierarchical clustering was applied to analyze the expression profiles of circRNAs of the GSE96964 database in OS cells (MG63, 143B, U2OS and HOS), human osteoblast cells (hFOB 1.19) and OS MTX-resistant cells (ZOS, ZOSM, and U2OS_MTX). (B) Levels of circRNAs were displayed in 3 groups. (C) Venn diagrams showed that there are 8 circRNAs among the three groups, and the levels of the 8 circRNAs are also shown. (D) Hsa_circ_0000073 expression was confirmed in cDNA and gDNA of U2OS and MG-63 cells using divergent primers and convergent primers. (E) The stability of hsa_circ_0000073 was determined in U2OS and MG-63 cells after treatment with RNase R by qRT-PCR assays. (F) Hsa_circ_0000073 expression was further determined using qRT-PCR analysis in hFOB 1.19 and OS cells (MG63, 143B, U2OS and HOS). (G) qRT-PCR analysis of hsa_circ_0000073 in OS and paired paratumor tissues. (H) qRT-PCR analysis of hsa_circ_0000073 in stage I-II and stage III-IV (UICC staging). (I) Kaplan-Meier analysis was applied to estimate the survival curve of hsa_circ_0000073 in OS. *P<0.05, **P<0.01.