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Drosha 敲低后宫颈癌中靶蛋白的蛋白质组学鉴定。

Proteomic identification of target proteins following Drosha knockdown in cervical cancer.

机构信息

Department of Obstetrics and Gynecology, Tongji Medical College, Union Hospital, Huazhong University of Science and Technology, Wuhan, Hubei, P.R. China.

出版信息

Oncol Rep. 2013 Nov;30(5):2229-37. doi: 10.3892/or.2013.2672. Epub 2013 Aug 20.

Abstract

The nuclear microRNA (miRNA) processing enzyme Drosha is upregulated in cervical cancer, and its overexpression is related to an invasive tumour phenotype. However, the mechanisms that underlie this effect remain poorly understood. The aim of this study was to identify the potential targets of Drosha in cervical cancer. Here, we demonstrated that Drosha knockdown (Drosha-KD) inhibited proliferation, colony formation and the migration of cervical cancer cells in vitro. A global upregulation of proteins in Drosha-KD cells was revealed by two-dimensional gel electrophoresis (2-DE). Eighteen proteins were identified by liquid chromatography and tandem mass spectrometry technology (LC-MS/MS) from 21 selected protein spots that exhibited significant alterations in Drosha-KD cells. The majority of the identified proteins have been previously associated with tumour formation. The downregulation of tubulin 5β in Drosha-KD cervical cancer cells was further confirmed by western blotting. Our results suggest that Drosha affects the biological activity of cervical cancer cells by regulating the expression of numerous tumour-associated proteins.

摘要

核微小 RNA(miRNA)加工酶 Drosha 在宫颈癌中上调,其过表达与侵袭性肿瘤表型有关。然而,其作用机制仍知之甚少。本研究旨在鉴定 Drosha 在宫颈癌中的潜在靶标。在此,我们证明 Drosha 敲低(Drosha-KD)抑制了宫颈癌细胞的体外增殖、集落形成和迁移。二维凝胶电泳(2-DE)显示 Drosha-KD 细胞中存在蛋白质的全局上调。通过液相色谱和串联质谱技术(LC-MS/MS)从 21 个差异明显的蛋白点中鉴定出 18 个蛋白。其中大多数已鉴定的蛋白与肿瘤形成有关。Drosha-KD 宫颈癌细胞中微管蛋白 5β 的下调进一步通过 Western blot 得到证实。我们的结果表明,Drosha 通过调节许多与肿瘤相关的蛋白的表达来影响宫颈癌细胞的生物学活性。

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