T1期声门区喉癌差异蛋白质组学的初步分析

Preliminary proteomic analysis of human serum from patients with T1 glottic laryngeal carcinoma

目的采用差异蛋白质组学分析声门区T1期喉癌患者术前与术后血清蛋白质组,寻找靶标蛋白。方法提取声门区T1期喉癌患者术前与术后血清蛋白质,二维凝胶电泳分离后,利用ImageMaster软件寻找差异蛋白点,通过质谱鉴定差异表达蛋白质,Western Blot检测其结果的表达。Elisa试剂盒检测患者及对照组血清丙二醛(MDA)和超氧化物岐化酶(SOD)表达水平。结果共鉴定T1期声门区喉癌患者术前与术后差异蛋白质12个,其中6个蛋白质在术前样本组上调表达,而与术后组差异无统计学意义(P>0.05)。术前组MDA表达水平高于对照组与术后组(P<0.01),SOD表达水平低于对照组(P<0.05)。结论氧化应激和免疫反应相关蛋白质在T1期声门区喉癌检测与治疗中具有潜在重要作用,差异蛋白质对于反应T1期声门区喉癌病理进程具有一定的研究意义。

Objective Different serum protein levels in preand post-operative T1 glottic laryngeal carcinoma patients were analyzed using proteomics methods to determine tumor biomarker proteins. Methods Comparative proteomic analysis of human sera between preand post-operative T1 glottic laryngeal carcinoma patients was performed using twodimensional （2D） gel electrophoresis, in combination with matrix-assisted laser desorption ionization time of flight mass spectrometry and database searching, to determine alternative proteins in T1 glottic laryngeal carcinoma patients. Western blot analysis, as well as the measurement of malondialdehyde （MDA） and superoxide dismutase （SOD） levels, was also performed. Results Image analysis of 2D gels indicated that 16 protein spots corresponding to 12 proteins exhibited significantly different expression levels between the two groups of T1 glottic laryngeal carcinoma patients. Six protein expression levels were upregulated. Western blot analysis confirmed the results of proteome analysis. MDA levels in the pretreatment group were higher than those in the control and post-treatment groups. By contrast, SOD levels in the pre-treatment group were lower than those in the control and post-treatment groups. Conclusion Proteomics results combined with bioinformatics analysis indicated the important roles of oxidant stress and immune response as targets in the monitoring, as well as treatment, of T1 glottic laryngeal carcinoma. The differentially expressed proteins may also reflect the pathological progress of T1 glottic laryngeal carcinoma and should be further studied.