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PRR11的下调通过抑制Wnt/β-连环蛋白信号通路影响骨肉瘤的增殖、迁移和侵袭。

Down-regulation of PRR11 affects the proliferation, migration and invasion of osteosarcoma by inhibiting the Wnt/β-catenin pathway.

作者信息

Li Ke, Yu Hongtao, Zhao Chunbing, Li Jing, Tan Rui, Chen Lei

机构信息

Orthopedic Center, First Affiliated Hospital, School of Medicine, Shihezi University, No.107 North 2nd Road, Shihezi, Xinjiang 832008, P. R. China.

Department of Pharmacy, First Affiliated Hospital, School of Medicine, Shihezi University, No.107 North 2nd Road, Shihezi, Xinjiang 832008, P. R. China.

出版信息

J Cancer. 2021 Sep 13;12(22):6656-6664. doi: 10.7150/jca.62491. eCollection 2021.

Abstract

This study aim to explore the effect of down-regulation of PRR11 (proline-rich protein 11) on the proliferation, invasion, migration, Wnt/β-catenin signaling pathway and EMT of osteosarcoma cells. Immunohistochemical staining, fluorescent quantitative PCR and western blotting were used to detect the expression level of PRR11 in osteosarcoma tissues and osteosarcoma cells. After SiRNA down-regulated the expression level of PRR11, CCK8 was used to detect cell proliferation ability, Transwell chamber to detect cell invasion ability, scratch test to detect cell migration ability, and flow cytometry to detect cell apoptosis. Western blotting was used to detect the expression levels of wnt/β-catenin pathway related proteins and key epithelial-mesenchymal transition proteins. PRR11 is highly expressed in osteosarcoma tissues, and its expression level is related to tumor size, Enneking stage of tumor, lymph node metastasis and patient prognosis. The low expression of PRR11 can inhibit the proliferation, migration and invasion of osteosarcoma cells, and promote apoptosis. Down-regulating the expression of PRR11 will inhibit the expression of Wnt pathway related proteins β-catenin and p-GSK-3β, enhance the expression of p-β-catenin, GSK-3β, and increase the expression of downstream genes CyclinD1 and c-Myc in the Wnt pathway. At the same time, the expression of PRR11 was down-regulated, the epithelial marker E-cadherin was significantly increased, and the expression levels of mesenchymal markers Vimentin and Fibronectin were significantly reduced. Down-regulation of PRR11 can inhibit the proliferation, migration and invasion of osteosarcoma cells, and its mechanism may be related to down-regulation of PRR11 to inhibit the Wnt/β-catenin signaling pathway and thus prevent the EMT process. Therefore, PRR11 may be used as an oncogene to promote the occurrence and development of osteosarcoma, and is a potential prognostic indicator and therapeutic target in osteosarcoma.

摘要

本研究旨在探讨下调富脯氨酸蛋白11(PRR11)对骨肉瘤细胞增殖、侵袭、迁移、Wnt/β-连环蛋白信号通路及上皮-间质转化(EMT)的影响。采用免疫组织化学染色、荧光定量PCR及蛋白质印迹法检测PRR11在骨肉瘤组织及骨肉瘤细胞中的表达水平。在小干扰RNA(SiRNA)下调PRR11表达水平后,采用细胞计数试剂盒8(CCK8)检测细胞增殖能力,Transwell小室检测细胞侵袭能力,划痕试验检测细胞迁移能力,流式细胞术检测细胞凋亡情况。采用蛋白质印迹法检测Wnt/β-连环蛋白通路相关蛋白及关键上皮-间质转化蛋白的表达水平。PRR11在骨肉瘤组织中高表达,其表达水平与肿瘤大小、肿瘤Enneking分期、淋巴结转移及患者预后相关。PRR11低表达可抑制骨肉瘤细胞的增殖、迁移和侵袭,并促进细胞凋亡。下调PRR11的表达会抑制Wnt通路相关蛋白β-连环蛋白和磷酸化糖原合成酶激酶-3β(p-GSK-3β)的表达,增强磷酸化β-连环蛋白(p-β-catenin)、糖原合成酶激酶-3β(GSK-3β)的表达,并增加Wnt通路下游基因细胞周期蛋白D1(CyclinD1)和原癌基因c-Myc的表达。同时,下调PRR11的表达后,上皮标志物E-钙黏蛋白显著增加,间质标志物波形蛋白和纤连蛋白的表达水平显著降低。下调PRR11可抑制骨肉瘤细胞的增殖、迁移和侵袭,其机制可能与下调PRR11抑制Wnt/β-连环蛋白信号通路从而阻止EMT进程有关。因此,PRR11可能作为癌基因促进骨肉瘤的发生发展,是骨肉瘤潜在的预后指标和治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e77/8518000/ecebadd5e74e/jcav12p6656g001.jpg

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