Overexpression of hsa_circ_0136666 predicts poor prognosis and initiates osteosarcoma tumorigenesis through miR-593-3p/ZEB2 pathway.

Chao Zhang,Kaizhen Yuan,Haibo Zhou,Chun Chen,Raoying Xie

doi:10.18632/aging.103273

Chao Zhang, Kaizhen Yuan + Show 3 more

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https://doi.org/10.18632/aging.103273

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Abstract

Background: Osteosarcoma (OS) is a type of malignant bone tumor with a growing incidence. Increasing studies indicate circular RNA (circRNA) has a vital function in tumorigenesis. Yet, how circRNA regulates OS development is not clear. In the present work, we aimed to investigate the roles of hsa_circ_0136666 in OS progression.Results: hsa_circ_0136666 was shown to be upregulated in OS and correlated with advanced stage and poor prognosis. Functional investigation using CCK8, colony formation assay and Transwell assay demonstrated that hsa_circ_0136666 promoted OS proliferation, migration and invasion, but inhibited cell death. Additionally, we identified hsa_circ_0136666 was a molecular sponge for miR-593-3p to facilitate ZEB2 expression. MiR-593-3p and ZEB2 were inversely expressed in OS tissues. And hsa_circ_0136666 exerts oncogenic roles in OS relying on miR-593-3p and ZEB2.Conclusion: Our results demonstrate the involvement of hsa_circ_0136666 in regulating OS tumorigenesis and it may be a therapeutic target.Methods: The expression of hsa_circ_0136666 was analyzed by qRT-PCR in OS tissues and cell lines. Proliferation was measured via CCK8 and colony formation assays. Migration and invasion were determined through Transwell assay. Luciferase reporter assay was utilized to determine the interaction between hsa_circ_0136666 and miR-593-3p or between miR-593-3p and ZEB2. Animal experiment was performed to investigate the role of hsa_circ_0136666 in vivo.

Highlights

Osteosarcoma (OS) is the most frequent and aggressive cancer in bone among young people [1]
We showed that hsa_circ_0136666 expression was upregulated in OS tissues and predicted poor prognosis
Based on Cell counting kit-8 (CCK8), Transwell and apoptosis detection assays, we found that miR-593-3p inhibition rescued the proliferation, migration, invasion and survival of U2OS and Saos2 cells transfected with hsa_circ_0136666 siRNA (Figure 4C–4F)

Summary

Introduction

Osteosarcoma (OS) is the most frequent and aggressive cancer in bone among young people [1]. It is urgently required to illustrate the molecular mechanism of OS progression and identify effective therapeutic targets for OS patients. A growing number of studies have indicated circRNAs have important functions involved in human diseases, including cancer [6, 7]. CircRNA circ_0026134 modulates cell growth and migration in non-small cell lung cancer (NSCLC) by inhibiting miR-1256 and miR-1287 [11]. CircRNA circ-FOXM1 regulates NSCLC proliferation, migration and invasion via repressing miR-1304 [12]. Functional investigation using CCK8, colony formation assay and Transwell assay demonstrated that hsa_circ_0136666 promoted OS proliferation, migration and invasion, but inhibited cell death. We identified hsa_circ_0136666 was a molecular sponge for miR-593-3p to facilitate ZEB2 expression. Conclusion: Our results demonstrate the involvement of hsa_circ_0136666 in regulating OS tumorigenesis and it may be a therapeutic target. Animal experiment was performed to investigate the role of hsa_circ_0136666 in vivo

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