Department of Biochemistry and Molecular Biology, Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China ; State Key Laboratory of Molecular Oncology, Beijing Key Laboratory for Carcinogenesis and Cancer Prevention, Cancer Institute (Hospital), Peking Union Medical College & Chinese Academy of Medical Sciences, P.O. Box 2258, Beijing 100021, China.
Department of Head and Neck Surgical Oncology, Cancer Institute (Hospital), Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100021, China.
Dis Markers. 2014;2014:594093. doi: 10.1155/2014/594093. Epub 2014 Feb 11.
This study was aimed to create a large-scale laryngeal cancer relevant secretory/releasing protein database and further discover candidate biomarkers.
Primary tissue cultures were established using tumor tissues and matched normal mucosal tissues collected from four laryngeal cancer patients. Serum-free conditioned medium (CM) samples were collected. These samples were then sequentially processed by SDS-PAGE separation, trypsin digestion, and LC-MS/MS analysis. The candidates in the database were validated by ELISA using plasma samples from laryngeal cancer patients, benign patients, and healthy individuals.
Combining MS data from the tumor tissues and normal tissues, 982 proteins were identified in total; extracellular proteins and cell surface proteins accounted for 15.0% and 4.3%, respectively. According to stringent criteria, 49 proteins were selected as candidates worthy of further validation. Of these, human tissue kallikrein 6 (KLK6) was verified. The level of KLK6 was significantly increased in the plasma samples from the cancer cohort compared to the benign and healthy cohorts and moreover showed a slight decrease in the postoperative plasma samples in comparison to the preoperative plasma samples.
This laryngeal cancer-derived protein database provides a promising repository of candidate blood biomarkers for laryngeal cancer. The diagnostic potential of KLK6 deserves further investigation.
本研究旨在创建一个大规模的喉癌相关分泌/释放蛋白数据库，并进一步发现候选生物标志物。
使用从 4 位喉癌患者收集的肿瘤组织和匹配的正常黏膜组织建立原代组织培养。收集无血清条件培养基 (CM) 样本。然后通过 SDS-PAGE 分离、胰蛋白酶消化和 LC-MS/MS 分析对这些样本进行连续处理。使用喉癌患者、良性患者和健康个体的血浆样本通过 ELISA 对数据库中的候选物进行验证。
将肿瘤组织和正常组织的 MS 数据结合起来，共鉴定出 982 种蛋白质；细胞外蛋白质和细胞表面蛋白质分别占 15.0%和 4.3%。根据严格的标准，选择了 49 种有进一步验证价值的候选蛋白。其中，人组织激肽释放酶 6（KLK6）得到了验证。与良性和健康队列相比，癌症队列的血浆样本中 KLK6 水平显著升高，与术前血浆样本相比，术后血浆样本略有下降。
该喉癌衍生蛋白数据库为喉癌提供了有前途的候选血液生物标志物库。KLK6 的诊断潜力值得进一步研究。
