KLF5‑induced miR‑487a augments the progression of osteosarcoma cells by targeting NKX3‑1 <em>in vitro</em>

Luo,Anyu; Liu,Hanlin; Huang,Chen

doi:10.3892/ol.2022.13378

KLF5‑induced miR‑487a augments the progression of osteosarcoma cells by targeting NKX3‑1 in vitro

Authors:
- Anyu Luo
- Hanlin Liu
- Chen Huang
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Affiliations: Department of Orthopedics, Hanyang Hospital Affiliated to Wuhan University of Science and Technology, Wuhan, Hubei 430051, P.R. China
Published online on: June 14, 2022 https://doi.org/10.3892/ol.2022.13378
Article Number: 258
Copyright: © Luo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

MicroRNAs (miRNAs or miRs) are involved in the development and progression of numerous types of cancer however their role in osteosarcoma has not been fully clarified. The present study aimed to use high‑throughput bioinformatics analysis as well as in vitro experiments to investigate the potential role of transcription factors, miRNAs and their targets in the progression of osteosarcoma. miRNA data and clinical information of osteosarcoma were obtained from Gene Expression Omnibus database to investigate differentially expressed miRNAs. The expression of miRNAs/mRNAs in osteosarcoma cell lines was detected via reverse transcription‑quantitative (RT‑qPCR). MTT and colony formation assay were used to determine cell proliferation ability and transwell assay was used to observe cell invasion and migration ability. A total of four prediction algorithms for miRNA‑mRNA interactions were used to determine potential target genes of miR‑487a. Predicted target genes were used to intersect with overlapped differentially expressed genes (DEGs) from GSE12865 and The Cancer Genome Atlas osteosarcoma datasets. Expression of NK3 homeobox 1 (NKX3‑1) was analyzed by western blotting and RT‑qPCR assay. Dual luciferase assay was conducted to verify whether NKX3‑1 was a direct target of miR‑487a. The regulatory association between Kruppel‑like factor 5 (KLF5) and miR‑487a was detected using chromatin immunoprecipitation assay. miR‑487a was upregulated in osteosarcoma tissue (GSE65071 and GSE28423) and cell lines (HOS and MG63). miR‑487a mimic promoted proliferation, migration and invasion of osteosarcoma cells. NKX3‑1 was a direct target of miR‑487a and transfection of NKX3‑1 plasmid reversed the effect of miR‑487a on proliferation, migration and invasion of osteosarcoma cells. KLF5 enhanced miR‑487a expression by directly binding to its promoter region and miR‑487a inhibitor reversed the effect of KLF5 on proliferation, migration and invasion of osteosarcoma cells. The present results indicated that KLF5/miR‑487a signaling promoted invasion and metastasis of osteosarcoma cells via targeting NKX3‑1.

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