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采用基质辅助激光解吸/电离飞行时间质谱分析法对口腔癌患者的唾液进行分析。

Analysis of the saliva from patients with oral cancer by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

作者信息

Chen Yu-Chie, Li Tzu-Ying, Tsai Ming-Fei

机构信息

Department of Applied Chemistry, National Chiao Tung University, Hsinchu 300, Taiwan.

出版信息

Rapid Commun Mass Spectrom. 2002;16(5):364-9. doi: 10.1002/rcm.588.

Abstract

Using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), this study analyzed the saliva obtained from patients with oral cancer and compared these mass spectra with those obtained from healthy controls. Saliva without pre-treatment was mixed directly with a sinapinic acid matrix. Alpha-amylase (57 kDa) dominated the high mass range in the MALDI mass spectra of the saliva from healthy subjects, but the peak was suppressed for patients with oral cancer and was replaced by a peak at m/z 66 k in the spectra of patients' samples (15 out of 20). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with in-gel tryptic digestion combined with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was employed to characterize this 66-kDa protein, which was thus shown to be albumin. However, based on SDS-PAGE results, concentrations of both alpha-amylase and albumin in patients' saliva were significantly higher than those in healthy subjects. This discrepancy was shown to be due to MALDI suppression effects due to the albumin. MALDI-MS thus has potential as a possible rapid diagnostic screening tool for oral cancer.

摘要

本研究使用基质辅助激光解吸/电离质谱(MALDI-MS)分析了口腔癌患者的唾液,并将这些质谱与健康对照者的质谱进行了比较。未经预处理的唾液直接与芥子酸基质混合。在健康受试者唾液的MALDI质谱中,α-淀粉酶(57 kDa)在高质量范围内占主导地位,但口腔癌患者的该峰被抑制,并且在患者样本的质谱图中(20例中有15例)被m/z 66 k处的峰所取代。采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)结合胶内胰蛋白酶消化与基质辅助激光解吸/电离飞行时间(MALDI-TOF)来表征这种66 kDa的蛋白质,结果表明其为白蛋白。然而,根据SDS-PAGE结果,患者唾液中α-淀粉酶和白蛋白的浓度均显著高于健康受试者。这种差异被证明是由于白蛋白的MALDI抑制作用所致。因此,MALDI-MS有潜力作为一种可能的口腔癌快速诊断筛查工具。

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