Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY 10032, USA.
Clin Chem Lab Med. 2010 Feb;48(2):271-8. doi: 10.1515/CCLM.2010.066.
There is a need to identify reliable markers for malignant mesothelioma. Soluble mesothelin related peptides (SMRP) and osteopontin (OPN) have gained interest in recent years for this purpose.
SMRP (Fujirebio Diagnostics Inc.) and OPN (R&D Inc.) ELISA methods were evaluated for multiple parameters. Concentrations were measured in blood from patients with mesothelioma, normal healthy volunteers, and patients with other (non-mesothelioma) cancers. In silico analysis was performed using the GeoProfiles database. At the protein level, SMRP and OPN were measured in cell culture supernatants, and values were compared in patients pre- and post-extrapleural pneumonectomy.
The SMRP assay demonstrates intra-assay CVs of 2.3% and 3% (at 4.6 nM and 13.7 nM, respectively), and inter-assay CVs of 3.5% and 3.7% at the same concentrations. The limit of detection (LOD) is 0.182 nM. The OPN assay demonstrates intra-assay CVs of 5.8%, 4.1%, and 5.2% (at 1.9, 5.1, and 11.1 ng/mL, respectively), and inter-assay CVs of 8.5%, 8.4%, and 12.1% at the same concentrations. The LOD is 0.032 ng/mL. Both SMRP and OPN in mesothelioma patients were significantly higher than in patients with other (non-mesothelioma) cancer and in healthy volunteers. The two markers do not appear to correlate with each other and exhibit different tissue expression patterns. Protein concentrations of these markers are highest in different sets of cell lines. Finally, SMRP but not OPN concentrations were decreased in five of seven consecutive patients after extrapleural pneumonectomy, compared to their respective pre-operative values.
These assays provide reliable and reproducible quantitation of SMRP and OPN proteins. Both are increased in mesothelioma patients compared to non-mesothelioma controls. However, the two analytes do not correlate with each other and show distinct expression profiles and protein expression. Concentrations of SMRP but not OPN are decreased in post-surgical samples. Our results further characterize these markers, establish assay performance characteristics, and lay the groundwork for further studies to measure these markers in blood.
需要确定恶性间皮瘤的可靠标志物。近年来，可溶性间皮素相关肽（SMRP）和骨桥蛋白（OPN）在这方面引起了关注。
评估了 SMRP（富士瑞必诊断公司）和 OPN（R&D 公司）ELISA 方法的多个参数。测量了间皮瘤患者、健康志愿者和其他（非间皮瘤）癌症患者的血液浓度。使用 GeoProfiles 数据库进行了计算机分析。在蛋白质水平上，测量了细胞培养上清液中的 SMRP 和 OPN 值，并比较了胸膜外肺切除术前后患者的值。
SMRP 检测法的组内 CV 分别为 2.3%和 3%（分别在 4.6 nM 和 13.7 nM 时），组间 CV 分别为 3.5%和 3.7%。检测限（LOD）为 0.182 nM。OPN 检测法的组内 CV 分别为 5.8%、4.1%和 5.2%（分别在 1.9、5.1 和 11.1 ng/mL 时），组间 CV 分别为 8.5%、8.4%和 12.1%。LOD 为 0.032 ng/mL。间皮瘤患者的 SMRP 和 OPN 均明显高于其他（非间皮瘤）癌症患者和健康志愿者。这两种标志物似乎彼此不相关，表现出不同的组织表达模式。这些标志物的蛋白浓度在不同的细胞系中最高。最后，与术前值相比，胸膜外肺切除术后五名连续患者中的 SMRP 但不是 OPN 浓度降低。
这些检测方法提供了 SMRP 和 OPN 蛋白的可靠和可重复定量。与非间皮瘤对照组相比，间皮瘤患者的这两种分析物均升高。然而，这两种分析物彼此不相关，并且表现出不同的表达谱和蛋白表达。手术样本中 SMRP 但不是 OPN 的浓度降低。我们的结果进一步描述了这些标志物，确定了检测方法的性能特征，并为进一步研究测量血液中的这些标志物奠定了基础。
