REG γ在骨肉瘤中的致癌作用是通过激活Wnt/β-连环蛋白信号通路来实现的。

The oncogenic role of REG γ is exerted by activating the Wnt/β-catenin signaling pathway in osteosarcoma.

作者信息

Yin Zhiqiang, Peng Zhibin, Wang Zhichao, Meng Qinggang

机构信息

Department of Orthopedics, The Fourth Affiliated Hospital of Harbin Medical University Harbin, Heilongjiang Province, China.

The Key Laboratory of Myocardial Ischemia, Chinese Ministry of Education Harbin, Heilongjiang Province, China.

出版信息

Am J Transl Res. 2020 Feb 15;12(2):563-573. eCollection 2020.

PMID:32194904

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7061849/

Abstract

BACKGROUND

Proteasome activator γ (REG γ) expression was found to be upregulated and to play critical roles in several cancers. However, the effect of REG γ on osteosarcoma (OS) remains unclear. The objective of the present study was to explore the clinical significance of REG γ and its function in regulating the progression of OS.

METHODS

Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting (WB) and immunohistochemistry (IHC) analyses were performed to detect the expression levels of REG γ in OS tissues and cell lines. Then, the effects of REG γ expression on OS cell behavior were analyzed by Cell Counting Kit-8 (CCK-8), ethylene deoxyuridine (EdU), colony formation, flow cytometry, wound healing and transwell assays. The protein and mRNA levels of components involved in the Wnt/β-catenin pathway were evaluated using WB and qRT-PCR, respectively.

RESULTS

We found that REG γ expression was significantly upregulated in both OS tissues and cell lines. Our assay results confirmed that knockdown of REG γ inhibited cell proliferation, migration, and invasion and induced apoptosis and cell cycle arrest in OS. Additionally, through WB and qRT-PCR analyses, we found that REG γ depletion markedly decreased the β-catenin, cyclin D1 and c-myc expression levels and increased the GSK-3β expression levels in OS cell lines.

CONCLUSIONS

Our results revealed that REG γ plays an oncogenic role in OS by activating the Wnt/β-catenin pathway, indicating that REG γ may be a promising therapeutic target for OS patients.

摘要

背景

蛋白酶体激活剂γ（REGγ）的表达在多种癌症中上调并发挥关键作用。然而，REGγ对骨肉瘤（OS）的影响仍不清楚。本研究的目的是探讨REGγ的临床意义及其在调节OS进展中的作用。

方法

采用定量逆转录-聚合酶链反应（qRT-PCR）、蛋白质印迹法（WB）和免疫组织化学（IHC）分析检测REGγ在OS组织和细胞系中的表达水平。然后，通过细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）、集落形成、流式细胞术、伤口愈合和Transwell实验分析REGγ表达对OS细胞行为的影响。分别使用WB和qRT-PCR评估Wnt/β-连环蛋白信号通路相关成分的蛋白质和mRNA水平。

结果

我们发现REGγ在OS组织和细胞系中的表达均显著上调。我们的实验结果证实，敲低REGγ可抑制OS细胞的增殖、迁移和侵袭，并诱导细胞凋亡和细胞周期停滞。此外，通过WB和qRT-PCR分析，我们发现敲低REGγ可显著降低OS细胞系中β-连环蛋白、细胞周期蛋白D1和c-myc的表达水平，并增加糖原合成酶激酶-3β（GSK-3β）的表达水平。