鼻咽癌高癌家系不同体质证型鼻咽癌患者鼻咽病灶组织蛋白双向凝胶电泳分析

Proteomic features of nasopharyngeal tissue sample of NPC familial patients in different constitutional patterns determined by 2-Dimensional electrophoresis

目的建立鼻咽癌高癌家系鼻咽癌患者气虚体质证型及湿热体质证型高分辨率双向凝胶电泳特征图谱,探讨其证本质相关性蛋白质组学特征。方法设鼻咽癌高癌家系鼻咽癌患者气虚体质组、湿热体质组、其他体质证型组,散发性鼻咽癌患者组,鼻咽癌高癌家系健康成员对照组,共计5组。分别获取鼻咽组织标本,提取总蛋白质,固相pH梯度双向凝胶电泳分离,经考马斯亮蓝显色后,Image Master 2D Elite4.01图像分析软件进行电泳结果分析。结果各组标本均获得分辨率高、重复性好的双向凝胶电泳图谱,平均蛋白质点数分别为860±20,854±25,842±30,867±20及857±15。比较分析,发现鼻咽癌高癌家系鼻咽癌患者气虚体质组与湿热体质组,差异蛋白质点10个。结论鼻咽癌高癌家系鼻咽癌患者不同体质证型蛋白质谱发生了明显变化,提示同病异证存在不同的本质内涵,为进一步研究鼻咽癌高癌家系鼻咽癌患者中医体质证型的本质内涵提供了实验依据。

Objective To identify the characteristics of high resolution 2-dimensional electrophoregrams of the total protein separated from nasopharyngeal focus tissues of cases with nasopharyngeal carcinoma （NPC） in either constitutional pattems of Qi deficiency or dampness-heat in familial NPC to explore its essential qualities of constitutional patterns in terms of proteomics. Methods Included in this study were 5 groups of eases in total, i.e. 3 groups of NPC eases with the constitutional pattern of Qi deficiency, dampnessheat and the mixed group of other patterns of constitution respectively, sampled from the familial lines so called as familial NPC, the group of cases with sporadic NPC in general population, and the healthy group of adults as controls, with tissue samples taken from their nasopharynx separately. Then, total protein was extracted from these groups of tissues in turn and separated by immobilized pH gradientbased 2-DE programs. Following the staining procedures to the gels by Coomassie brilliant blue G-250, the differentially expressed protein dots were identified by Image Master 2D Elite4.01 analysis software. Results Well-resolved and reproducible 2-DE image patterns of studied nasopharyngeal tissue samples were obtained and the average protein dots were 860±20, 854±25, 842±30, 867±20 and 857±15 respectively for these 5 groups of experimental cases. As the results of comparative analysis among these groups, ten differential protein dots were revealed out from the familial NPC groups of NPC eases with the constitutional pattern either Qi deficiency or dampness-heat. Conclusions Significantly changed characteristics in the expressive spectrogram of proteins has taken place among the NPC patients with different constitutional patterns in familial NPC, suggesting that cases with different syndromes but in the same disease may show different proteomic features, which should provide helpful information in further study on the essential qualities of different constitutional patterns in familial NPC.