relationships linking amplification level to gene over-expression in gliomas连接放大在神经胶质瘤基因表达水平的关系.pdfVIP

Relationships Linking Amplification Level to Gene Over-Expression in Gliomas Nicolas Vogt1,2,3, Anne Gibaud 1,2,3, Anna Almeida1,2,3¤, Isabelle Ourliac-Garnier 1,2,3, Michelle Debatisse1,2,3, Bernard Malfoy1,2,3* 1 Institut Curie, Centre de Recherche, Paris, France, 2 CNRS UMR3244, Paris, France, 3 UPMC, Paris, France Abstract Background: Gene amplification is thought to promote over-expression of genes favouring tumour development. Because amplified regions are usually megabase-long, amplification often concerns numerous syntenic or non-syntenic genes, among which only a subset is over-expressed. The rationale for these differences remains poorly understood. Methodology/Principal Finding: To address this question, we used quantitative RT-PCR to determine the expression level of a series of co-amplified genes in five xenografted and one fresh human gliomas. These gliomas were chosen because we have previously characterised in detail the genetic content of their amplicons. In all the cases, the amplified sequences lie on extra-chromosomal DNA molecules, as commonly observed in gliomas. We show here that genes transcribed in non- amplified gliomas are over-expressed when amplified, roughly in proportion to their copy number, while non-expressed genes remain inactive. When specific antibodies were available, we also compared protein expression in amplified and non- amplified tumours. We found that protein accumulation barely correlates with the level of mRNA expression in some of these tumours. Conclusions/Significance: Here we show that the tissue-specific pattern of gene expression is maintained upon amplification in gliomas. Our study relies on a single type of tumour and a limited number of cases. However, it strongly suggests that, even when amplified, genes that are normally silent in a given cell type p