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UHRF1通过以Rb1依赖的方式下调E-钙黏蛋白的表达来促进人骨肉瘤细胞的侵袭。

UHRF1 promotes human osteosarcoma cell invasion by downregulating the expression of E‑cadherin in an Rb1‑dependent manner.

作者信息

Liu Wei, Qiao Rui Hong, Wang Dong Ming, Huang Xiao Wei, Li Bing, Wang Dong

机构信息

Department of Orthopedics, Second Clinical Medical College, Shanxi Medical University, Taiyuan, Shanxi 215006, P.R. China.

Department of Gastroenterology, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, Hubei 226001, P.R. China.

出版信息

Mol Med Rep. 2016 Jan;13(1):315-20. doi: 10.3892/mmr.2015.4515. Epub 2015 Nov 6.

Abstract

Ubiquitin‑like with plant homeodomain (PHD) and RING‑finger domain 1 (UHRF1) maintains methylation patterns following DNA replication and is expressed at high levels in various types of human cancer. UHRF1 has been identified as a novel oncogene involved in the pathogenesis of hepatocellular carcinoma. Previous studies have demonstrated that inhibition of the expression of UHRF1 suppresses the proliferation of cancer cells. However, the role of UHRF1 in human osteosarcoma has not been investigated. The present study examined the expression levels of UHRF1 and retinoblastoma 1 (Rb1) in human osteosarcoma cell lines by western blot analysis. Stable overexpression of UHRF1 or knockdown of Rb1 was achieved by lentiviral transfection. Subsequently, a Cell Counting Kit-8 assay and a cell invasion assay were performed to detect the biological functions of UHRF1 in vitro. The results of the present study demonstrated that UHRF1 promoted the proliferation of human osteosarcoma cells. The present study also reported that UHRF1 was able to enhance the invasion of osteosarcoma cells in a retinoblastoma 1 (Rb1)‑dependent manner. UHRF1 promoted invasion in Rb1‑positive osteosarcoma cells, but not in Saos‑2 cells with homozygous loss of Rb1. Similarly, knockdown of Rb1 in Rb1‑positive osteosarcoma cells enhanced levels of invasion and eliminated the regulation of invasion by UHRF1. UHRF1 was found to inhibit the mRNA and protein expression levels of Rb1. Furthermore, deletion of Rb1 was found to suppress the expression of E‑cadherin and promote epithelial‑to‑mesenchymal transition (EMT). In addition, the overexpression of UHRF1 inhibited the expression of E‑cadherin and promoted EMT via the suppression of Rb1. These data demonstrated that UHRF1 promotes osteosarcoma cell invasion by downregulating the expression of E‑cadherin and increasing EMT in an Rb1‑dependent manner.

摘要

含植物同源结构域(PHD)和泛素样结构域以及RING指结构域1(UHRF1)可在DNA复制后维持甲基化模式,并在多种人类癌症中高表达。UHRF1已被鉴定为一种参与肝细胞癌发病机制的新型癌基因。先前的研究表明,抑制UHRF1的表达可抑制癌细胞的增殖。然而,UHRF1在人类骨肉瘤中的作用尚未得到研究。本研究通过蛋白质免疫印迹分析检测了UHRF1和视网膜母细胞瘤1(Rb1)在人类骨肉瘤细胞系中的表达水平。通过慢病毒转染实现了UHRF1的稳定过表达或Rb1的敲低。随后,进行细胞计数试剂盒-8检测和细胞侵袭检测以体外检测UHRF1的生物学功能。本研究结果表明,UHRF1促进人类骨肉瘤细胞的增殖。本研究还报道,UHRF1能够以视网膜母细胞瘤1(Rb1)依赖的方式增强骨肉瘤细胞的侵袭能力。UHRF1促进Rb1阳性骨肉瘤细胞的侵袭,但对Rb1纯合缺失的Saos-2细胞无此作用。同样,在Rb1阳性骨肉瘤细胞中敲低Rb1可增强侵袭水平,并消除UHRF1对侵袭的调节作用。研究发现,UHRF1可抑制Rb1的mRNA和蛋白表达水平。此外,发现Rb1的缺失可抑制E-钙黏蛋白的表达并促进上皮-间质转化(EMT)。此外,UHRF1的过表达通过抑制Rb1抑制E-钙黏蛋白的表达并促进EMT。这些数据表明,UHRF1通过下调E-钙黏蛋白的表达并以Rb1依赖的方式增加EMT来促进骨肉瘤细胞的侵袭。

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