Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455, USA.
Electrophoresis. 2010 Jan;31(4):599-610. doi: 10.1002/elps.200900441.
Ovarian cancer is the fifth leading cause of cancer death for women in the US, yet survival rates are over 90% when it is diagnosed at an early stage, highlighting the need for biomarkers for early detection. To enhance the discovery of tumor-specific proteins that could represent novel serum biomarkers for ovarian cancer, we depleted serum of highly abundant proteins which can mask the detection of proteins present in serum at low concentrations. Three commercial immunoaffinity columns were used in parallel to deplete the highly abundant proteins in serum from 60 patients with serous ovarian carcinoma and 60 non-cancer controls. Medium and low abundance serum proteins from each serum pool were then evaluated by the quantitative proteomic technique of differential in-gel electrophoresis. The number of protein spots that were elevated in ovarian cancer sera by at least twofold ranged from 36 to 248, depending upon the depletion and separation methods. From the 33 spots picked for MS analysis, nine different proteins were identified, including the novel candidate ovarian cancer biomarkers leucine-rich alpha2 glycoprotein-1 and ficolin 3. Western blotting validated the relative increases in serum protein levels for three of the proteins identified, demonstrating the utility of this approach for the identification of novel serum biomarkers for ovarian cancer.
卵巢癌是美国女性癌症死亡的第五大主要原因，但如果在早期诊断，存活率超过 90%，这突显了对早期检测生物标志物的需求。为了增强对肿瘤特异性蛋白的发现，这些蛋白可能代表卵巢癌的新型血清生物标志物，我们耗尽了血清中高丰度的蛋白，这些蛋白可以掩盖低浓度存在于血清中的蛋白的检测。我们平行使用了三种商业免疫亲和柱来耗尽来自 60 名浆液性卵巢癌患者和 60 名非癌症对照者的血清中的高丰度蛋白。然后，通过差异凝胶电泳的定量蛋白质组学技术评估每个血清池的中低丰度血清蛋白。根据耗尽和分离方法的不同，卵巢癌血清中至少升高两倍的蛋白点数量从 36 个到 248 个不等。从 33 个用于 MS 分析的斑点中，鉴定出了 9 种不同的蛋白质，包括新型候选卵巢癌生物标志物富含亮氨酸的α2 糖蛋白 1 和纤维胶凝素 3。Western blotting 验证了三种鉴定出的蛋白质的血清蛋白水平的相对增加，证明了这种方法对鉴定卵巢癌新型血清生物标志物的实用性。
