Research Paper Volume 14, Issue 10 pp 4486—4499

Long noncoding RNA KCNQ1OT1 inhibits osteoclast differentiation by regulating the miR-128-3p/NFAT5 axis

Hengshuo Zhang1,2, *, , Lu Chen1,2, *, , Ziyu Wang1,2, , Zhenqian Sun1,2, , Yu Shan1,2, , Qinghui Li1,2, , Linzeng Qi1,2, , Hongliang Wang1, , Yunzhen Chen1, ,

  • 1 Department of Orthopedics, Qilu Hospital of Shandong University, Jinan, Shandong 250012, PR China
  • 2 Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250012, PR China
* Co-first author

Received: January 22, 2022       Accepted: May 7, 2022       Published: May 19, 2022
How to Cite

Copyright: © 2022 Zhang 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.


Noncoding RNAs play an important role in regulating osteoclast differentiation. We investigated whether and how potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1 (KCNQ1OT1), a long noncoding RNA, regulates osteoclast differentiation. We found that the expression of KCNQ1OT1 was downregulated in osteoporotic bone tissue. Then transfection of KCNQ1OT1 overexpression vectors or small interfering RNAs showed that the proliferation, migration, and osteoclast differentiation of RAW 264.7 cells were inhibited by KCNQ1OT1 upregulation, while they were promoted by KCNQ1OT1 knockdown. Interestingly, we found and confirmed that miR-128-3p was a target of KCNQ1OT1 using online databases, dual luciferase reporter assays and quantitative real-time polymerase chain reaction, and that it inhibited the expression of miR-128-3p. Moreover, we confirmed that miR-128-3p directly targeted nuclear factor of activated T cell 5 (NFAT5), a protein that combines with osteoprotegerin and thus regulates osteoclastogenesis with the presence of the receptor activator of nuclear factor κB ligand. Furthermore, we demonstrated that both the knockdown of KCNQ1OT1 and the overexpression of miR-128-3p attenuate the expression of NFAT5, while upregulating the osteoclastogenesis markers c-Fos, NFATc1, and Ctsk. The results from overexpression of KCNQ1OT1 and the inhibition of miR-128-3p were contrary to the above. Finally, we found that the inhibition of osteoclast differentiation by KCNQ1OT1 overexpression could be rescued using a miR-128-3p mimic, while the enhancement of migration and osteoclast differentiation by si-NFAT5 could be reversed with a miR-128-3p inhibitor. These results suggested that KCNQ1OT1 regulates the osteoclast differentiation via the miR-128-3p/NFAT5 axis.


ANOVA: analysis of variance; BMD: bone mineral density; ceRNAs: competing endogenous RNAs; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hBMSCs: human bone marrow mesenchymal stem cells; KCNQ1OT1: potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1; lncRNA: long noncoding RNA; miRNA: microRNA; mMSCs: mouse mesenchymal stem cells; MUT: mutant-type; ncRNAs: noncoding RNAs; NFAT5: nuclear factor of activated T cell 5; NC: negative control; OP: osteoporosis; OPG: osteoprotegerin; PVDF: polyvinylidenedifluoride; qRT-PCR: quantitative real-time polymerase chain reaction; RANKL: receptor activator of nuclear factor κB ligand; RIPA: radio immunoprecipitation assay; siRNA: small interfering RNA; SD: standard deviation; TFs: transcription factors; TRAP: Tartrate-resistant acid phosphatase; WT: wild type.