基于“同病异治”的大肠癌不同证候血清蛋白组学探索研究  

Experimental study in serum proteomics on different syndromes of colorectal cancer in the adjuvant therapy

作　　者：徐玉芬[1] 杨新妹[1] 孙校男[2] 郭勇[3,4] 杨维泓[3,4] 

机构地区：[1]浙江省嘉兴市第一医院肿瘤科,314000 [2]浙江中医药大学,310053 [3]浙江中医药大学附属第一医院 [4]浙江省中医院肿瘤科,310006

出　　处：《中华全科医学》2015年第4期647-649,663,F0003,共5页Chinese Journal of General Practice

基　　金：浙江省科技厅公益技术应用研究项目(2010-C33140);浙江省中医药科学研究基金(2011ZA031);2014年浙江省嘉兴市重点学科(肿瘤科)建设项目[嘉市卫发(2014)72号]

摘　　要：目的利用双向凝胶电泳和质谱分析对辅助治疗期大肠癌脾气虚证、胃阴虚证、平和证型患者血清蛋白质组进行分析,探索同一疾病不同证候的血清蛋白质表达差异。方法选取辅助治疗期大肠癌脾气虚证、胃阴虚证患者各8例,以大肠癌平和证型患者6例作为对照组,采集清晨空腹血清标本,通过亲和层析法去除血清高丰度白蛋白和免疫球蛋白,运用Cy Dye荧光染料进行标记,并设立内标,上样于同一胶中进行双向凝胶电泳分离,电泳结束后分别用488 nm、530 nm、633 nm波长激光激发扫描获得不同样品的蛋白质组图谱,所得蛋白质图谱应用De Cyder 6.5软件进行分析,选择蛋白质表达水平上升或下降120%以上的差异蛋白质进行质谱鉴定及生物信息学分析。结果在大肠癌辅助治疗期不同证候中筛选出13个表达有显著差异的蛋白质,其中7个蛋白质被确定:血红素前体、α-1-B-糖蛋白、色氨酸羟化酶-1、结合珠蛋白在胃阴虚证、脾气虚证患者血清中表达均上调,而富含亮氨酸的α-2-糖蛋白-1表达均下调。胰蛋白酶抑制剂HI30、转甲状腺素蛋白在胃阴虚证患者中表达上调,但在脾气虚证患者中表达无显著差异。结论大肠癌辅助治疗期脾气虚证、胃阴虚证形成存在不同的分子基础,可以作为辨证论治的潜在客观指标,胃阴虚证的产生可能与胰蛋白酶抑制剂HI30、转甲状腺素蛋白上调有关。Objective Serum protein among colorectal cancer patients with stomach-yin deficiency, spleen-qi deficiency and single symptom were studied by two-dimensional gel electrophoresis and mass spectrometry in the period of adjuvant therapy. Methods There were three groups：8 patients with stomach-yin deficiency,8 patients with spleen-qi deficiency and 6 patients with single symptom. High abundance serum albumin and immunoglobulin of the samples were removed by affinity chromatography and then labeled with fluorescent dyes, and analyzed by 2D gel electrophoresis. The gels were respectively imaged by Typhoon 9400 using different emission filter and spot-features were analyzed by DeCyder Image Quant TM V6.5 software, and the different proteins were identified with mass spectrum. Results Twelve different pro- teins were selected in different syndromes and 7 of them were identified. The expression of hemopexin precursor, alpha-1- B-glycoprote,Tryptophan hydroxylase 1 and haptoglobin were upregulation, whereas Leucine-rich alpha-2-glycoproteinl was decreased in both groups of patients with stomach-yin deficiency and spleen-qi deficiency. Trypsin inhibitor HI30 and Transthyretin expression were upregulated only in the patients with stomach-yin deficiency. Conclusion Different syn- dromes of colorectal cancer patients were in different molecular basis, which could be used as the potential objective marks. And stomach yin deficiency may result from the trypsin inhibitor HI30 and transthyretin upregulation.

关 键 词：大肠癌 脾气虚证 胃阴虚证 血清蛋白质组学 

分 类 号：R735.34[医药卫生—肿瘤] R242[医药卫生—临床医学]