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[iTRAQ] 胶质瘤-iTRAQ

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发表于 2015-12-28 17:19:11 | 显示全部楼层 |阅读模式

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Cancer Res. 2009 Nov 15;69(22):8545-54. doi: 10.1158/0008-5472.CAN-09-1778. Epub 2009 Nov 10.
Loss of collapsin response mediator Protein1, as detected by iTRAQ analysis, promotes invasion of humangliomas expressing mutant EGFRvIII.
IF=
14年9.329
13年9.284
12年8.65
11年7.856
研究关键词 胶质瘤 iTRAQ
研究概要:
      多形性胶质母细胞瘤(GBM)是最常见且恶性程度最高的中枢神经系统原发恶性肿瘤,在GBM的诸多遗传学改变中,EGFR的突变(EGFRvIII)是最常见的,具有EGFRvIII的GBM通常具有更高的增值和侵袭活性,本研究目的在于探讨EGFRvIII促进GBM增值侵袭的分子机制。研究包括3部分:1。利用4标的iTRAQ技术检测野生型EGFR(wtEGFR)(2标)和EGFRvIII(2标)的GBM组织的蛋白表达差异,所得数据进行生物信息学分析后,发现CRMP1在表达EFGRvIII的GBM组织中表达显著下调;2.扩大组织样本量后,用Wertern blot、免疫组化、和qRT-PCR分别在蛋白水平和RNA水平验证VRMP1在表达EFGRvIII的组织中低表达;3.利用GBM细胞系U87和U373下调CRMP1的表达水平,观察细胞表型变化,验证CRMP1功能。本研究获得结论:CEMP1在表达EGFRvIII的GBM中表达降低,增加肿瘤的侵袭力。
Abstract
Glioblastoma multiforme (GBM) is the most common and lethal primary human brain tumor. GBMs are characterized by a variety of genetic alterations, among which oncogenic mutations of epidermal growth factor receptor (EGFRvIII) is most common. GBMs harboring EGFRvIII have increased proliferation and invasive characteristics versus those expressing wild-type (wt) EGFR. To identify the molecular basis of this increased tumorgenic phenotype, we used iTRAQ-labeling differential proteomic analysis. Among several differentially expressed proteins, we selected CRMP1, a protein implicated in cellular invasion that was markedly decreased in GBMs expressing EGFRvIII, for further study. The differential expression of CRMP1 was confirmed in a panel of human GBM cell lines and operative specimens that express wtEGFR or mutant EGFRvIII by quantitative real-time PCR, Western blot, and immunohistochemical analysis. In human GBM samples, decreased expression of CRMP1 correlated with EGFRvIIIpositivity. Knockdown of CRMP1 by siRNA resulted in increased invasion of wtEGFR expressing human GBM cells (U87 and U373) to those found in isogenic GBM cells. Exogenous expression of EGFRvIII in these wtEGFR-expressing GBM cells promoted their ability to invade and was accompanied by decreased expression of CRMP1. Rescuing CRMP1 expression decreased invasion of the EGFRvIII-expressing GBM cells by tilting the balance between Rac and Rho. Collectively, these results show that the loss of CRMP1 contribute to the increased invasive phenotype of humanGBMs expressing mutant EGFRvIII.

PMID: 19903856
免费全文链接
http://cancerres.aacrjournals.org/content/69/22/8545.long









Cancer Res-2009-IF8.65.pdf

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