Research Paper Volume 13, Issue 13 pp 17274—17284

Circ_0081001 down-regulates miR-494-3p to enhance BACH1 expression and promotes osteosarcoma progression

Shizhang Liu1, , Keke Duan1, , Xiaoxia Zhang1, , Xiane Cao2, , Xixia Wang3, , Fanbin Meng2, , Huitong Liu1, , Bingqiang Xu1, , Xi Wang1, ,

  • 1 Department of Orthopedics, Shaanxi Provincial People’s Hospital, Xian 710068, Shanxi, China
  • 2 Department of Orthopedics, Linyi People's Hospital, Linyi 276100, Shandong, China
  • 3 Department of Central Sterile Supply, Linyi People's Hospital, Linyi 276100, Shandong, China

Received: September 1, 2020       Accepted: May 24, 2021       Published: June 30, 2021
How to Cite

Copyright: © 2021 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


The study was aimed at deciphering the function and mechanism of circ_0081001 in osteosarcoma (OS). In this study, quantitative real-time polymerase chain reaction (qRT-PCR) was utilized for quantifying circ_0081001, miR-494-3p, and BTB domain and CNC homolog 1 (BACH1) mRNA expressions in OS tissues and cells. Cell counting kit-8 (CCK-8) assay, together with 5-Ethynyl-2'-deoxyuridine (EdU) assay, was performed for evaluating cell proliferation; the alterations in apoptosis were analyzed utilizing flow cytometry; Transwell assay was conducted for examining cell migration and invasion; moreover, Western blot was utilized for the quantification of BACH1 protein expression; bioinformatics, dual-luciferase reporter gene, and RNA-binding protein immunoprecipitation assays were executed for validating the binding relationships between circ_0081001 and miR-494-3p, and between miR-494-3p and BACH1. As shown, circ_0081001, whose expression was elevated in OS tissues, had a negative association with miR-494-3p expression and a positive correlation with BACH1 expression. After circ_0081001 was overexpressed, the proliferation, migration, and invasion of OS cells were boosted but the apoptosis was reduced, whereas miR-494-3p exhibited opposite effects. The binding sites between circ_0081001 and miR-494-3p, and between miR-494-3p and the 3’UTR of BACH1 were experimentally verified. In conclusion, circ_0081001/miR-494-3p/BACH1 axis promoted the malignant biological behaviors of OS cells.


OS: osteosarcoma; circRNAs: circular RNAs; miRNAs: microRNAs; ceRNA: competitive endogenous RNA; BACH1: BTB domain and CNC homolog 1; CCK-8: cell counting kit-8; EdU: 5-Ethynyl-2'-deoxyuridine; qRT-PCR: quantitative real-time polymerase chain reaction; RIP: RNA-binding protein immunoprecipitation.