Ma Yanlei, Peng Jiayuan, Huang Long, Liu Weijie, Zhang Peng, Qin Huanlong
Department of Surgery, The Sixth People's Hospital Affiliated to Shanghai Jiao Tong University, Shanghai, PR China.
Electrophoresis. 2009 Aug;30(15):2591-9. doi: 10.1002/elps.200900082.
Identification of specific protein markers for colorectal cancer (CRC) could provide a basis for its early diagnosis and detection, as well as clues to the molecular mechanisms governing cancer progression. In the present study, 2-D DIGE coupled with MS was used to screen for biomarker candidates in the serum proteome of ten human CRC samples and ten healthy control samples. After pooling identical amounts of serum proteins (based on total protein concentration), albumin/IgG was depleted under partially denaturing conditions. Subsequently, the serum samples were labeled with three different CyDyes, and separated by 2-D DIGE. After analysis with the biological variation analysis module of the DeCyder software, only three spots were found to be significantly elevated in all patient groups (with ratios from 1.52 to 9.08), whereas five spots were significantly down-regulated in patients (with ratios from -1.23 to -10.21) (t-test; p<0.05). Finally, two potential biomarkers, Transaldolase 1 and thyroid receptor interactor, were chosen for validation and analysis by ELISA with the serum of 30 CRC patients and 30 healthy controls. The serum levels of the two proteins correlated well with the 2-D DIGE results. Thus, 2-D DIGE approaches show great promise for biomarker discovery in CRC.
鉴定结直肠癌（CRC）的特异性蛋白质标志物可为其早期诊断和检测提供依据，同时也能为了解癌症进展的分子机制提供线索。在本研究中，二维差异凝胶电泳（2-D DIGE）结合质谱（MS）用于筛选10例人类CRC样本和10例健康对照样本血清蛋白质组中的潜在生物标志物。在基于总蛋白浓度合并等量血清蛋白后，在部分变性条件下去除白蛋白/免疫球蛋白（IgG）。随后，血清样本用三种不同的CyDye进行标记，并通过二维差异凝胶电泳进行分离。使用DeCyder软件的生物学变异分析模块进行分析后，发现只有三个斑点在所有患者组中显著升高（比值为1.52至9.08），而五个斑点在患者中显著下调（比值为-1.23至-10.21）（t检验；p<0.05）。最后，选择转醛醇酶1和甲状腺受体相互作用分子这两种潜在生物标志物，用30例CRC患者和30例健康对照的血清进行酶联免疫吸附测定（ELISA）验证和分析。这两种蛋白质的血清水平与二维差异凝胶电泳结果高度相关。因此，二维差异凝胶电泳方法在CRC生物标志物发现方面显示出巨大潜力。
