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目的:对口腔黏膜癌变的蛋白组学分析和分子标志物筛选可能为早期诊断、预防和分子治疗提供依据和帮助。方法:对3例石蜡包埋口腔黏膜癌变组织样本,利用激光显微切割技术捕获口腔黏膜癌变上皮和癌旁正常黏膜上皮,抽提组织样本蛋白,利用高效液相色谱-质谱联用进行口腔黏膜癌变组织的蛋白组学分析。结果:激光显微切割-高效液相色谱质谱联用可以分析400~700个多肽序列,鉴定出100~200个蛋白,涉及到细胞骨架、细胞代谢、信号转导、细胞周期等各个方面。结合文献,对其中的角蛋白表达变化进行了分析和验证。结论:显微切割结合高效液相色谱质谱可以实现对石蜡包埋组织的蛋白组学研究,具有较高的重复性和可靠性。角蛋白表达谱的变化验证了该平台的作用和价值,也可作为口腔黏膜癌变的潜在标志物。
OBJECTIVE: The proteomic analysis and molecular marker screening of oral mucosal carcinogenesis may provide evidence and help for early diagnosis, prevention and molecular therapy. Methods: Three cases of paraffin-embedded cancerous tissue samples were paraffin-embedded, and laser-microdissected technique was used to capture the epithelial and para-normal mucosal epithelia of oral mucosa. Protein extracted from the tissue samples was extracted by high performance liquid chromatography-mass spectrometry Proteomic analysis of cancerous tissues. Results: Laser microdissection - high performance liquid chromatography-mass spectrometry (LC-MS / MS) could analyze 400-700 polypeptide sequences and identify 100-200 protein involved in various aspects such as cytoskeleton, cell metabolism, signal transduction, cell cycle and so on. Combined with the literature, the change of keratin expression was analyzed and verified. Conclusions: Microdissection combined with high performance liquid chromatography-mass spectrometry can achieve the proteomics study of paraffin-embedded tissues with high repeatability and reliability. Changes in keratin expression profiles validate the role and value of this platform and may also serve as potential markers of carcinogenesis in oral mucosa.