Sarkissian Gaiane, Fergelot Patricia, Lamy Pierre-Jean, Patard Jean-Jacques, Culine Stephane, Jouin Patrick, Rioux-Leclercq Nathalie, Darbouret Bruno
Research and Development Department, Cezanne SAS, Parc Scientifique Georges Besse, Nimes, France.
Clin Chem. 2008 Mar;54(3):574-81. doi: 10.1373/clinchem.2007.090837. Epub 2008 Jan 17.
No validated renal cell carcinoma (RCC) marker is known for detection of asymptomatic disease in selected populations or for prognostic purposes or treatment monitoring. We identified immunogenic proteins as tumor markers for RCC by combining conventional proteome analysis with serological screening, and we investigated the diagnostic clinical value of such markers in serum.
We studied the immunogenic protein expression profile of CAL 54, a human RCC cell line, by 2-dimensional electrophoresis combined with immunoblotting using sera from healthy donors compared with RCC patients. We developed a homogeneous, fluorescent, dual-monoclonal immunoassay for metalloproteinase 7 (MMP-7) and used it to measure MMP-7 in sera from 30 healthy donors, 30 RCC patients, and 40 control patients.
Pro-MMP-7 (29 kDa; pI 7.7) in the CAL 54 cell line secretome was an immunogenic protein reactive with RCC patient sera but not with control sera. The concentrations of pro-MMP-7 were increased (P <0.0001) in sera of RCC patients (median 7.56 microg/L; range 3.12-30.5 microg/L) compared with healthy controls (median 2.13 microg/L; range 0.17-3.5 microg/L). Serum pro-MMP-7 had a sensitivity of 93% (95% CI 78%-99%) at a specificity of 75% (59%-87%) for RCC in the samples tested.
Proteomics technology combined with serology led to the identification of serum pro-MMP-7 as a marker of RCC and represents a powerful tool in searching for candidate proteins as biomarkers.
目前尚无经过验证的肾细胞癌（RCC）标志物可用于在特定人群中检测无症状疾病、进行预后评估或治疗监测。我们通过将传统蛋白质组分析与血清学筛查相结合，确定了免疫原性蛋白作为RCC的肿瘤标志物，并研究了此类标志物在血清中的诊断临床价值。
我们采用二维电泳结合免疫印迹法，利用健康供体与RCC患者的血清，研究了人RCC细胞系CAL 54的免疫原性蛋白表达谱。我们开发了一种用于金属蛋白酶7（MMP-7）的均相、荧光、双单克隆免疫测定法，并用于检测30名健康供体、30名RCC患者和40名对照患者血清中的MMP-7。
CAL 54细胞系分泌组中的前MMP-7（29 kDa；pI 7.7）是一种免疫原性蛋白，与RCC患者血清反应，但与对照血清不反应。与健康对照（中位数2.13μg/L；范围0.17 - 3.5μg/L）相比，RCC患者血清中前MMP-7的浓度升高（P <0.0001）（中位数7.56μg/L；范围3.12 - 30.5μg/L）。在测试样本中，血清前MMP-7对RCC的敏感性为93%（95%CI 78% - 99%），特异性为75%（59% - 87%）。
蛋白质组学技术与血清学相结合，导致血清前MMP-7被鉴定为RCC的标志物，并代表了一种寻找候选蛋白作为生物标志物的强大工具。
