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肌球蛋白重链 10(MYH10)基因沉默通过抑制 Wnt/β-连环蛋白通路减少胶质瘤细胞系 U251、T98G 和 SHG44 的细胞迁移和侵袭。

Myosin Heavy Chain 10 (MYH10) Gene Silencing Reduces Cell Migration and Invasion in the Glioma Cell Lines U251, T98G, and SHG44 by Inhibiting the Wnt/β-Catenin Pathway.

机构信息

Department of Neurosurgery, 2nd Ward, Taihe Hospital, Shiyan, Hubei, China (mainland).

Department of Orthopedic Surgery, 3rd Ward, Taihe Hospital, Shiyan, Hubei, China (mainland).

出版信息

Med Sci Monit. 2018 Dec 15;24:9110-9119. doi: 10.12659/MSM.911523.

DOI:10.12659/MSM.911523
PMID:30552850
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6319164/
Abstract

BACKGROUND The myosin heavy chain 10 or MYH10 gene encodes non-muscle myosin II B (NM IIB), and is involved in tumor cell migration, invasion, extracellular matrix (ECM) production, and epithelial-mesenchymal transition (EMT). This study aimed to investigate the effects of the MYH10 gene on normal human glial cells and glioma cell lines in vitro, by gene silencing, and to determine the signaling pathways involved. MATERIAL AND METHODS The normal human glial cell line HEB, and the glioma cell lines, U251, T98G, and SHG44 were studied. Plasmid transfection silenced the MYH10 gene. The cell counting kit-8 (CCK-8) assay evaluated cell viability. Cell migration and invasion were evaluated using scratch and transwell assays. Western blot measured the protein expression levels, and quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression levels, for MYH10, metastasis-associated protein 1 (MTA-1), matrix metalloproteinase (MMP)-1, MMP-9, tissue inhibitor of metalloproteinases 2 (TIMP2), collagen 1, E-cadherin, vimentin, Wnt3a, β-catenin, and cyclin D1. RESULTS The MYH10 gene was overexpressed in U251, T98G, and SHG44 cells. MYH10 expression was down-regulated following siMYH10 plasmid interference, which also inhibited glioma cell migration and invasion. MYH10 gene silencing resulted in reduced expression of MTA-1, MPP-2, MMP-9 and vimentin, and increased expression of TIMP-2, E-cadherin and collagen 1 at the protein and mRNA level, and inhibited the Wnt/β-catenin pathway. CONCLUSIONS In human glioma cell lines, silencing the MYH10 gene reduced cell migration and invasion, by inhibiting the Wnt/β-catenin pathway, which may regulate the ECM and inhibit EMT in human glioma.

摘要

背景

肌球蛋白重链 10 或 MYH10 基因编码非肌肉肌球蛋白 II B(NM IIB),参与肿瘤细胞迁移、侵袭、细胞外基质(ECM)产生和上皮-间充质转化(EMT)。本研究旨在通过基因沉默研究 MYH10 基因对体外正常人神经胶质细胞和神经胶质瘤细胞系的影响,并确定相关信号通路。

材料和方法

研究了正常人神经胶质细胞系 HEB 和神经胶质瘤细胞系 U251、T98G 和 SHG44。质粒转染沉默 MYH10 基因。细胞计数试剂盒-8(CCK-8)测定细胞活力。划痕和 Transwell 测定评估细胞迁移和侵袭。Western blot 测定蛋白表达水平,实时定量聚合酶链反应(qRT-PCR)检测 MYH10、转移相关蛋白 1(MTA-1)、基质金属蛋白酶(MMP)-1、MMP-9、金属蛋白酶组织抑制剂 2(TIMP2)、胶原 1、E-钙黏蛋白、波形蛋白、Wnt3a、β-连环蛋白和细胞周期蛋白 D1 的 mRNA 表达水平。

结果

U251、T98G 和 SHG44 细胞中 MYH10 基因过表达。siMYH10 质粒干扰后 MYH10 表达下调,抑制神经胶质瘤细胞迁移和侵袭。MYH10 基因沉默导致 MTA-1、MMP-2、MMP-9 和波形蛋白的表达减少,TIMP-2、E-钙黏蛋白和胶原 1 的表达增加,蛋白和 mRNA 水平抑制 Wnt/β-连环蛋白通路。

结论

在人神经胶质瘤细胞系中,沉默 MYH10 基因通过抑制 Wnt/β-连环蛋白通路减少细胞迁移和侵袭,可能调节 ECM 并抑制人神经胶质瘤中的 EMT。

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