肾透明细胞癌血清差异性蛋白质表达及临床意义研究

The expression of serum differentiated protein in patients with renal clear cell carcinoma and its clinical significance

目的 研究旨在利用蛋白组学技术,通过检测肾透明细胞癌患者和健康人群的蛋白表达水平差异、肾透明细胞癌患者手术前后蛋白表达水平的差异,寻找生物标志物.方法 选取2012年1月至2016年12月就诊于本院诊断为肾透明细胞癌的患者88例作为实验组,肾囊肿患者和泌尿系结石患者120例作为对照组,对实验组的患者在术后进行随访,留取实验组患者术前、后每次随访的血清,留取对照组患者的血清,使用表面增强激光解吸离子化飞行时间质谱（SELDI-TOF-MS）和CM10弱阳离子交换蛋白芯片技术系统检测差异性蛋白质,并用ZUCI-PDAS软件进行分析.结果 检测出质荷比为15949、9306、5915的三种蛋白,对应Bcl-2家族细胞凋亡调节蛋白、Krueppel样因子8、血清淀粉样蛋白4,对照组与实验组相比、实验组术前和术后相比,蛋白质表达水平均有统计学差异（P＜0.05）,这三种蛋白作为生物标记物,预测肾透明细胞癌的准确度为91.3％（190/208）,敏感度为89.8％（79/88）,特异度为92.5％（111/120）,此外笔者在既往发表的肾透明细胞癌研究所使用的基因芯片数据（Affymetrix HG-U133a和HG-U133b）中进一步验证所筛选的三个基因,发现与正常组织相比较,癌组织中上述三中基因的表达量表达量差异均有统计学意义,且表达量与肿瘤的分级相关.结论 利用SELDI-TOF-MS技术检测差异性表达的蛋白质是可行的,Bcl-2家族细胞凋亡调节蛋白、Krueppel样因子8、血清淡粉样蛋白4为本研究中筛选出的肾透明细胞癌血清肿瘤标记物.

 Objective To explore the tumor marker using proteomics techniques and find the serum differentiated protein in patients with renal clear cell carcinoma through the comparison between renal clear cell carcinoma patients and healthy patients,and between the serum protein level before and after surgery.Methods We enrolled 88 patients with renal clear cell carcinoma as the experimental group,120 patients with cystonephrosis or urinary calculus as the control group.Participants＇serum were collected.We followed up the patients in the experimental group after the surgery and collected their blood sample each time.The surface enhanced laser desorption/ionization time of flight mass spectrometry and CM10 weak cation exchange protein chip technique were used to analyze the differentiated protein expression,the ZUCI-PDAS software was used to analyze.Results Three protein with M/Z 15949,9306,and 5915 were selected as potential biological marker,they were Bcl-2 inter-acting mediator of cell death,ubiquitous kruppel like factor,and serum amyloid protein 4.Compared with the control group and postoperatively sample,the protein level was significantly different （P 〈0.05）.The accuracy,sensitivity,and specificity of the three protein in diagnosing renal cell carcinoma were 91.3% （190/208）,89.8% （79/88）,（111/120）,respectively.Conclusions It is applicable to use surface enhanced laser desorption/ionization time of flight mass spectrometry to explore the differentiated protein,Bcl-2 interacting mediator of cell death,ubiquitous kruppel like factor,and serum amyloid protein 4 are the three potential biological tumor marker in renal clear cell carcinoma.