College of Pharmacy, Chongqing Medical University, Chongqing 400016, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Dec 15;878(32):3395-401. doi: 10.1016/j.jchromb.2010.10.026. Epub 2010 Nov 4.
Urine is one of the most attractive analyte used for clinical diagnosis. NSCLC (non-small cell lung carcinoma), which includes adenocarcinoma, squamous cell carcinoma and large-cell carcinoma, is a leading cause of cancer-related deaths. In the present study, urinary proteomes of normal individuals and NSCLC patients were compared using 1D SDS-PAGE. From the distinctly differentially expressed bands in SDS-PAGE gel, 40 proteins were identified by chip-HPLC-MS/MS, including five proteins relevant to NSCLC. One of the selected proteins, alpha-1-antichymotrypsin (AACT), was further validated in urine by western blot and in lung tissue by immunohistochemistry staining. Higher expression level of AACT in NSCLC patients was observed by western blot when compared with normal urine samples. Significantly, the NSCLC tumor tissue (18 out of 20 cases, 90%) showed a significantly higher expression level of AACT compared to adjacent non-tumor lung tissue (3 out of 20 cases, 15%). These results establish AACT as a potential biomarker for objective and non-invasive diagnosis of NSCLC in urine and the other four NSCLC-related proteins were also listed.
尿液是用于临床诊断的最有吸引力的分析物之一。非小细胞肺癌（NSCLC）包括腺癌、鳞状细胞癌和大细胞癌，是癌症相关死亡的主要原因。在本研究中，使用 1D SDS-PAGE 比较了正常个体和 NSCLC 患者的尿液蛋白质组。通过芯片-HPLC-MS/MS 鉴定了 SDS-PAGE 凝胶中差异明显的表达带中的 40 种蛋白质，其中包括 5 种与 NSCLC 相关的蛋白质。所选蛋白质之一，α-1-抗糜蛋白酶（AACT），通过 Western blot 在尿液中进一步验证，通过免疫组织化学染色在肺组织中验证。与正常尿液样本相比，Western blot 观察到 NSCLC 患者的 AACT 表达水平更高。重要的是，与相邻非肿瘤肺组织（20 例中的 3 例，15%）相比，NSCLC 肿瘤组织（20 例中的 18 例，90%）中 AACT 的表达水平显著更高。这些结果确立了 AACT 作为尿液中 NSCLC 客观和非侵入性诊断的潜在生物标志物，列出了另外四种与 NSCLC 相关的蛋白质。
