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miR-301b在骨肉瘤中的过表达模式及其通过调控SNX10与骨肉瘤细胞行为的相关性

Overexpression Pattern of miR-301b in Osteosarcoma and Its Relevance with Osteosarcoma Cellular Behaviors via Modulating SNX10.

作者信息

Wang Yaozong, Sun Naikun, Zhang Zheyi, Zhou Yuanyuan, Liu Hongyi, Zhou Xu, Zhang Ying, Zhao Yilin

机构信息

Department of Orthopedics, Zhongshan Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, 361000, China.

Department of Orthopedics, The First Affiliated Hospital, School of Medicine, Xiamen University, Xiamen, 361000, China.

出版信息

Biochem Genet. 2023 Feb;61(1):87-100. doi: 10.1007/s10528-022-10241-4. Epub 2022 Jun 22.

Abstract

Prior studies have noted the importance of microRNAs (miRNAs) in development and progression of osteosarcoma (OS), but the influence of miR-301b is less investigated. This investigation aimed to explore the biological role of miR-301b/SNX10 in OS. GSE28423 and GSE28424 arrays delivered the corresponding miR-301b and sorting nexin 10 (SNX10) expression levels in OS samples. miR-301b and SNX10 expressions were also measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting in cells. Cell counting kit (CCK)-8 and transwell analysis were applied to measure cell characteristics. Luciferase reporter assay and Pearson correlation analysis were used to detect the relevance between miR-301b and SNX10. miR-301b was extremely increased in OS tissues compared with normal tissues, while SNX10 was decreased. The proliferation, invasion, and migration capabilities were limited following a low expression level of miR-301b whereas miR-301b overexpression promoted cellular malignant behaviors. miR-301b negatively targeted SNX10. The elevated SNX10 expression highlighted the inhibitory function on cell proliferation, migration, and invasion in OS cells treated by miR-301b inhibitor. Reduction of miR-301b induced the decrease of epithelial-mesenchymal transition (EMT)-related markers including N-cadherin, Vimentin, and matrix metallo-proteinase 9 (MMP)9. These results are added to the complete expanding field of the potential effects of miR-301b in OS cell malignant behaviors and demonstrate its promising role for further use to treat human OS.

摘要

先前的研究已经指出微小RNA(miRNA)在骨肉瘤(OS)发生发展中的重要性,但对miR-301b的影响研究较少。本研究旨在探讨miR-301b/SNX10在骨肉瘤中的生物学作用。GSE28423和GSE28424基因芯片检测了骨肉瘤样本中相应的miR-301b和分选连接蛋白10(SNX10)的表达水平。还通过定量逆转录-聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法检测了细胞中miR-301b和SNX10的表达。应用细胞计数试剂盒(CCK)-8和Transwell分析检测细胞特性。荧光素酶报告基因检测和Pearson相关性分析用于检测miR-301b与SNX10之间的相关性。与正常组织相比,骨肉瘤组织中miR-301b显著上调,而SNX10下调。miR-301b低表达时,细胞的增殖、侵袭和迁移能力受到限制,而miR-301b过表达则促进细胞的恶性行为。miR-301b对SNX10具有负向靶向作用。SNX10表达升高凸显了其对miR-301b抑制剂处理的骨肉瘤细胞增殖、迁移和侵袭的抑制作用。miR-301b的减少导致上皮-间质转化(EMT)相关标志物包括N-钙黏蛋白、波形蛋白和基质金属蛋白酶9(MMP)9的表达降低。这些结果进一步拓展了miR-301b在骨肉瘤细胞恶性行为中潜在作用的研究领域,并证明了其在治疗人类骨肉瘤方面的潜在应用价值。

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