UKM Medical Molecular Biology Institute (UMBI), Universiti Kebangsaan Malaysia, Level 7, Clinical Block, UKM Medical Centre, Jalan Yaacob Latiff, Bandar Tun Razak, 56000, Cheras, Kuala Lumpur, Malaysia.
Pathol Oncol Res. 2014 Jan;20(1):119-29. doi: 10.1007/s12253-013-9670-9. Epub 2013 Aug 8.
This study aimed to identify candidate proteins which may serve as potential biological markers for cervical cancer using 2D-DIGE. Serum samples of controls, patients with cervical intraepithelial neoplasia grade 3 (CIN 3), squamous cell carcinoma of early (SCC I and II) and late (SCC III and IV) stage were subjected to 2D-DIGE. Differentially expressed spots were identified by tandem mass spectrometry. Validation of candidate proteins in serum and tissue samples were then performed by ELISA and immunohistochemistry (IHC) analysis respectively. A total of 20 differentially expressed proteins were identified. These proteins were found to play key roles in the apoptosis pathway, complement system, various types of transportation such as hormones, fatty acids, lipid, vitamin E and drug transportation, coagulation cascade, regulation of iron and immunologic response. Based on their functional relevancy to the progression of various cancers, 4 proteins namely the complement factor H, CD5-like antigen, gelsolin and ceruloplasmin were chosen for further validation using ELISA. Biological network analysis showed that ceruloplasmin and gelsolin are closely interacted with the oncogene NF-κb. These two proteins were further validated using the IHC. Gelsolin and ceruloplasmin may serve as potential predictive biomarkers for the progression of high grade lesions.
本研究旨在通过 2D-DIGE 技术鉴定候选蛋白，这些蛋白可能成为宫颈癌潜在的生物标志物。收集对照组、宫颈上皮内瘤变 3 级（CIN3）、早期（SCC I 和 II）和晚期（SCC III 和 IV）鳞状细胞癌患者的血清样本进行 2D-DIGE。通过串联质谱鉴定差异表达的斑点。然后通过 ELISA 和免疫组织化学（IHC）分析分别对候选蛋白在血清和组织样本中的表达进行验证。共鉴定出 20 个差异表达蛋白。这些蛋白在细胞凋亡途径、补体系统、各种类型的运输（如激素、脂肪酸、脂质、维生素 E 和药物运输）、凝血级联反应、铁的调节和免疫反应中发挥关键作用。基于它们与各种癌症进展的功能相关性，选择了 4 种蛋白（补体因子 H、CD5 样抗原、凝溶胶蛋白和铜蓝蛋白）进行进一步的 ELISA 验证。生物网络分析显示，铜蓝蛋白和凝溶胶蛋白与癌基因 NF-κb 密切相互作用。使用 IHC 进一步验证了这两种蛋白。凝溶胶蛋白和铜蓝蛋白可能是高级别病变进展的潜在预测生物标志物。
