Knockdown of lncRNA C5orf66‑AS1 inhibits osteosarcoma cell proliferation and invasion via miR‑149‑5p upregulation

Zhang,Hui; Song,Jie

doi:10.3892/ol.2021.13018

Knockdown of lncRNA C5orf66‑AS1 inhibits osteosarcoma cell proliferation and invasion via miR‑149‑5p upregulation

Authors:
- Hui Zhang
- Jie Song
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Affiliations: Department of Orthopedics, Huangshi Central Hospital, Affiliated Hospital of Hubei Polytechnic University, Edong Healthcare Group, Huangshi, Hubei 435000, P.R. China, Department of Geriatrics, Huangshi Central Hospital, Affiliated Hospital of Hubei Polytechnic University, Edong Healthcare Group, Huangshi, Hubei 435000, P.R. China
Published online on: September 2, 2021 https://doi.org/10.3892/ol.2021.13018
Article Number: 757
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteosarcoma (OS) is the most common primary malignant bone tumor in the pediatric age group. Despite the various potential treatments for OS, the cure rate of patients with OS remains very low. An increasing number of long non‑coding RNAs (lncRNAs) have been identified as key regulators of the progression of malignant human tumors. However, the biological functions of the lncRNA C5orf66‑antisense 1 (C5orf66‑AS1) in OS are yet to be fully elucidated. The present study aimed to investigate the functions and underlying mechanisms of C5orf66‑AS1 in OS tissues and cell lines. Expression levels of C5orf66‑AS1 and microRNA (miRNA/miR)‑149‑5p in tissues from patients with OS and OS cells lines were evaluated using reverse transcription quantitative (RT‑q)PCR. The miRNA target interaction between C5orf66‑AS1 and miR‑149‑5p was predicted and verified using StarBase and dual‑luciferase reporter assays. Cell viability, migration, invasion and apoptosis were analyzed using Cell Counting Kit‑8, Transwell assays and ﬂow cytometry, respectively. In addition, the expression levels of migration‑ and apoptosis‑associated proteins [matrix metalloproteinase‑9 (MMP‑9), Bcl‑2 and Bax] were determined using western blotting and RT‑qPCR. The results demonstrated that C5orf66‑AS1 was significantly upregulated and miR‑149‑5p was significantly downregulated in OS tissues and cells (MG63 and U2OS). Bioinformatics analysis further confirmed that miR‑149‑5p could directly bind to C5orf66‑AS1. Furthermore, it was revealed that C5orf66‑AS1 negatively regulated the expression of miR‑149‑5p in OS cells, as confirmed by the inhibition of C5orf66‑AS1 expression and miR‑149‑5p upregulation in cells transfected with small interfering (si RNA targeting C5orf66‑AS1. In addition, C5orf66‑AS1 silencing significantly inhibited the proliferation, invasion and migration of U2OS cells, and stimulated cell apoptosis. These findings were reversed using miR‑149‑5p inhibitor. Increased Bax expression and decreased Bcl‑2 and MMP‑9 expression were also observed in C5orf66‑AS1‑siRNA transfected U2OS cells, compared with the control group. In summary, the results from the present study indicated that C5orf66‑AS1 knockdown inhibits OS cell proliferation and invasion via the upregulation of miR‑149‑5p. This findings may provide a promising novel target for the treatment of OS.

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