Department of Surgery, Division of Transplantation and Hepatobiliary Surgery, Erasmus University Medical Center, Rotterdam, The Netherlands.
PLoS One. 2013 Aug 16;8(8):e70918. doi: 10.1371/journal.pone.0070918. eCollection 2013.
For both patients and the outpatient clinic the frequent follow-up visits after a resection of colorectal cancer (CRC) are time consuming and due to large patient numbers expensive. Therefore it is important to develop an effective non-invasive test for the detection of colorectal liver metastasis (CRLM) which could be used outside the hospital. The urine proteome is known to provide detailed information for monitoring changes in the physiology of humans. Urine collection is non-invasive and urine naturally occurring peptides (NOPs) have the advantage of being easily accessible without labour-intensive sample preparation. These advantages make it potentially useful for a quick and reliable application in clinical settings. In this study, we will focus on the identification and validation of urine NOPs to discriminate patients with CRLM from healthy controls.
Urine samples were collected from 24 patients with CRLM and 25 healthy controls. In the first part of the study, samples were measured with a nano liquid chromatography (LC) system (Thermo Fisher Scientific, Germaring, Germany) coupled on-line to a hybrid linear ion trap/Orbitrap mass spectrometer (LTQ-Orbitrap-XL, Thermo Fisher Scientific, Bremen, Germany). A discovery set was used to construct the model and consecutively the validation set, being independent from the discovery set, to check the acquired model. From the peptides which were selected, multiple reaction monitoring (MRM's) were developed on a UPLC-MS/MS system.
Seven peptides were selected and applied in a discriminant analysis a sensitivity of 84.6% and a specificity of 92.3% were established (Canonical correlation:0.797, Eigenvalue:1.744, F:4.49, p:0.005). The peptides AGPP(-OH)GEAGKP(-OH)GEQGVP(-OH)GDLGA P(-OH)GP and KGNSGEP(-OH)GAPGSKGDTGAKGEP(-OH)GPVG were selected for further quantitative analysis which showed a sensitivity of 88% and a specificity of 88%.
Urine proteomic analysis revealed two very promising peptides, both part from collagen type 1, AGPP(-OH)GEAGKP(-OH)GEQGVP(-OH)GDLGAP(-OH)GP and KGNSGEP(-OH)GAPGSKGDTGAKGEP(-OH)GPVG which could detect CRLM in a non-invasive manner.
对于患者和门诊来说，结直肠癌（CRC）切除术后频繁的随访既耗时又昂贵，因为患者人数众多。因此，开发一种有效的非侵入性检测方法来检测结直肠肝转移（CRLM）非常重要，这种方法可以在医院外使用。尿液蛋白质组学可以为监测人体生理变化提供详细信息。尿液采集是非侵入性的，尿液中天然存在的肽（NOP）具有无需费力的样本制备即可轻松获取的优势。这些优势使其在临床环境中具有快速可靠应用的潜力。在这项研究中，我们将专注于鉴定和验证尿液 NOPs，以区分 CRLM 患者和健康对照者。
收集了 24 名 CRLM 患者和 25 名健康对照者的尿液样本。在研究的第一部分，使用纳升液相色谱（LC）系统（Thermo Fisher Scientific，Germaring，德国）与混合线性离子阱/轨道阱质谱仪（LTQ-Orbitrap-XL，Thermo Fisher Scientific，不来梅，德国）在线耦合进行测量。发现集用于构建模型，然后是验证集，与发现集独立，以检查获得的模型。从选择的肽中，开发了多个反应监测（MRM）在 UPLC-MS/MS 系统上进行。
选择了 7 个肽，并应用于判别分析，建立了 84.6%的灵敏度和 92.3%的特异性（典范相关：0.797，特征值：1.744，F：4.49，p：0.005）。肽 AGPP(-OH)GEAGKP(-OH)GEQGVP(-OH)GDLGA P(-OH)GP 和 KGNSGEP(-OH)GAPGSKGDTGAKGEP(-OH)GPVG 被选用于进一步的定量分析，其灵敏度为 88%，特异性为 88%。
尿液蛋白质组学分析显示了两种非常有前途的肽，均来自胶原蛋白 1，AGPP(-OH)GEAGKP(-OH)GEQGVP(-OH)GDLGAP(-OH)GP 和 KGNSGEP(-OH)GAPGSKGDTGAKGEP(-OH)GPVG，可通过非侵入性方式检测 CRLM。
