实验案例 为了开发新的临床免疫组化生物标记来区分肝细胞癌(sHCC)和发育异常的结节(DN),采用iTRAQ技术来筛选区分癌前病变和肝细胞癌的免疫组化生物标记。 Ø 已发表文献(IF5.12) iTRAQ-2DLC-ESI-MS/MSBased Identification of a New Set of Immunohistochemical Biomarkers for Classification of Dysplastic Nodules and Small Hepatocellular Carcinoma. J. Proteome Res. 2011, 10, 3418–3428 Ø 实验设计 样品取自LC,DN和sHCC三个不同的组织来源。提取蛋白后,分别用115,116,117来标记,混合蛋白并经过SCX柱液相分离后,zei后进入液质系统进行蛋白质谱鉴定。 Ø 结果 共有1951个蛋白被定量,在1.25倍p<0.05的显著差异下得到52个上调蛋白和95个下调蛋白。 挑选出来的候选生物标记进一步通过Western和免疫组化进行验证。进而用ROC曲线和逻辑回归模型来评估生物标记的诊断效果。
zei后ACY1和SQSTM1被选为新的生物标记,而所建立的逻辑回归模型包含ACY1,SQSTM1和CD34。这个模型的敏感性和特异性分别是96.1%和96.7%。 部分iTRAQ技术已发表文献 iTRAQ plus 18O: A new technique for target glycoprotein analysis , Talanta, Volume 91, 15 March 2012, Pages 122-127(方法学) Identification of transaldolaseas a novel serum biomarker for hepatocellular carcinoma metastasis using xenografted mouse modeland clinic samples , Cancer Letters 313 (2011) 154–166 (IF 4.8,血清样本) iTRAQ2DLCESIMS/MS Based Identification of a New Set ofimmunohistochemical Biomarkers forClassification of Dysplastic Nodules and SmallHepatocellular Carcinoma,J. Proteome Res. 2011, 10, 3418–3428(IF5.12,组织样本) Analysis of Transcriptional Factors and Regulation Networks in Patients with Acute Renal Allograft RejectionJournal of Proteome Research 2011, 10, 175–181(IF5.12,血清样本) Changes in proteomics profile during maturation of marrow-derived dendritic cells treated with oxidized low-density lipoprotein Proteomics 2011, 11, 1893–190(IF 4.42,细胞样本) LC-MS/MS Analysis of Ovarian Cancer Metastasis-Related Proteins Using a Nude Mouse Model: 14-3-3 Zeta as a Candidate BiomarkerJ. Proteome Res., 2010, 6180–6190(IF5.12,囊液样本)