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Quantitative proteome analysis of ovarian cancer tissues using a iTRAQ approach

机译:使用iTRAQ方法对卵巢癌组织进行蛋白质组定量分析

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摘要

Quantitative proteomics can be used as a screening tool for identification of differentially expressed proteins as potential biomarkers for cancers. Here, we comparatively analyzed the proteome profiles of ovarian cancer tissues and normal ovarian epithelial tissues. Using the high-throughput proteomic technology of isobaric tags for relative and absolute quantitation (iTRAQ)-coupled with two-dimensional-liquid chromatography-tandem mass spectrometry, 1,259 unique proteins were identified. Of those, 205 were potentially differentially expressed between ovarian cancer and normal ovarian tissues. Several of the potentially differentially expressed proteins were validated by Western blotting and real-time quantitative RT-PCR analyses. Furthermore, up-regulation of KRT8, PPA1, IDH2, and S100A11 were validated in ovarian tissue microarrays by immunohistochemistry. Silencing of S100A11 expression suppressed the migration and invasion properties of ovarian cancer cells in vitro. Our study represents the successful application of iTRAQ technology to an investigation of ovarian cancer. Many of the potentially differentially expressed proteins identified had not been linked to ovarian cancer before, and provide valuable novel insights into the underlying mechanisms of carcinogenesis in human ovarian cancer.
机译:定量蛋白质组学可以用作筛选工具,以鉴定差异表达的蛋白质作为癌症的潜在生物标记。在这里,我们比较分析了卵巢癌组织和正常卵巢上皮组织的蛋白质组图谱。使用等压标签的高通量蛋白质组学技术进行相对定量和绝对定量(iTRAQ),并与二维液相色谱-串联质谱联用,鉴定出1,259种独特蛋白质。其中,有205个在卵巢癌和正常卵巢组织之间可能存在差异表达。通过蛋白质印迹和实时定量RT-PCR分析验证了几种潜在差异表达的蛋白质。此外,通过免疫组织化学在卵巢组织微阵列中验证了KRT8,PPA1,IDH2和S100A11的上调。 S100A11表达的沉默抑制了体外卵巢癌细胞的迁移和侵袭特性。我们的研究代表了iTRAQ技术在卵巢癌研究中的成功应用。鉴定出的许多可能差异表达的蛋白质以前从未与卵巢癌相关,并为人卵巢癌致癌的潜在机制提供了有价值的新颖见解。

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