Department of Otorhinolaryngology, Head and Neck Surgery, Medical University of Vienna, A-1090 Vienna, Austria.
Department of Molecular Biotechnology, University of Applied Science Vienna, Helmut-Qualtinger-Gasse 2, A-1090 Vienna, Austria.
EPMA J. 2014 Nov 28;5(1):20. doi: 10.1186/1878-5085-5-20. eCollection 2014.
The analysis of biomarkers in saliva as a clinical application offers an attractive, simple and rapid diagnostic tool for the short- and long-term monitoring of pathological disorders and drug therapy. The collection of saliva, either in the pure or in its fractionated form, is a relatively easy and non-invasive procedure that is not harmful to the patients and has no complications at all. However, the fluid collection must be clearly defined due to variations in saliva composition, flow rate and day-to-day variability. In order to minimise possible variations, saliva from five patients without squamous cell carcinoma (SCC) pathology and five with suspicion of oral squamous carcinoma (OSCC) were collected and matched at different days and analysed by two-dimensional polyacrylamide gel electrophoresis (2DE-PAGE). Approximately 800 spots were identified, corresponding to 151 different gene products. The list of identified proteins includes a large number of structural proteins like keratins, keratin subunits, enzymes and enzyme inhibitors, cytokines, immunoglobulins as well as amylase and other salivary specific glycoproteins. The majority of proteins that are localised in oral epithelia cells were found as unsolved debris in saliva. One of the identified proteins was significantly overexpressed in OSCC and was selected for further validation by Western blot analysis.
唾液生物标志物分析作为一种临床应用,为短期和长期监测病理紊乱和药物治疗提供了一种有吸引力、简单和快速的诊断工具。唾液的采集,无论是纯唾液还是其分级形式,都是一种相对容易且非侵入性的程序,对患者无害,完全没有并发症。然而,由于唾液成分、流速和日常变化的差异,必须明确界定液体采集。为了最小化可能的变化,收集了 5 名无鳞状细胞癌(SCC)病理和 5 名疑似口腔鳞状细胞癌(OSCC)的患者的唾液,并在不同的日子进行匹配,然后通过二维聚丙烯酰胺凝胶电泳(2DE-PAGE)进行分析。鉴定出了大约 800 个斑点,对应于 151 种不同的基因产物。鉴定出的蛋白质列表包括大量的结构蛋白,如角蛋白、角蛋白亚基、酶和酶抑制剂、细胞因子、免疫球蛋白以及淀粉酶和其他唾液特异性糖蛋白。大多数定位于口腔上皮细胞的蛋白质被发现是唾液中的未解决碎片。在 OSCC 中,一种鉴定出的蛋白质显著过表达,并被选择用于 Western blot 分析进行进一步验证。