Profilin 1 is a Potential Biomarker for Bladder Cancer Aggressiveness
Profilin 1 is a Potential Biomarker for Bladder Cancer Aggressiveness
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DOI：
10.1074/mcp.m111.009449
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发表时间：
2012-04-01
影响因子：
7
通讯作者：
Vlahou, Antonia
中科院分区：
文献类型：
--
作者：
Zoidakis, Jerome;Makridakis, Manousos;Vlahou, Antonia
膀胱癌（BC）管理的最重要的临床需求是识别疾病侵袭性的生物标志物。尿液是生物标志物发现的“金矿”，然而，由于多种蛋白质含量低，尿液蛋白质组学变得具有挑战性。在本研究中，我们应用了一种基于使用固定化金属亲和色谱的尿蛋白分级策略，用于发现侵袭性BC的生物标志物。非侵入性患者的尿液样本（两个游泳池）和侵入性（两个池）BC经受固定化金属亲和色谱分级分离，并通过1D-SDS-PAGE、条带切除和液相色谱串联MS分析洗脱的蛋白质。在鉴定的蛋白质中，多个对应于对金属具有亲和性的蛋白质和/或报道被磷酸化的蛋白质，并且包括与BC具有证明的关联的蛋白质，例如MMP 9，纤维蛋白原形式和丛生蛋白。与固定化金属亲和层析结果一致，通过Western印迹或Elisa分析，进一步发现氨肽酶N、profilin 1和成髓细胞蛋白在侵袭性患者与非侵袭性BC和良性对照相比的尿液中差异表达，但表现出较高的个体间变异性。通过组织微阵列分析，发现profilin 1在侵袭性（T2+）与高风险非侵袭性（T1 G3）肿瘤的上皮细胞中表达显著降低，偶尔在基质中表达;重要的是，这种模式与不良预后和死亡率增加密切相关。在T24 BC细胞中研究了profilin 1的功能相关性，其中通过使用抗体阻断蛋白质导致细胞运动性降低，伴随着肌动蛋白聚合的降低。总的来说，我们的研究涉及尿蛋白的分级方法的应用，并且作为该分析的一个主要结果，揭示了profilin 1与BC的关联，为其在BC分层中的进一步研究铺平了道路。Molecular & Cellular Proteomics 11：10.1074/mcp.M111.009449，1-15，2012.
Of the most important clinical needs for bladder cancer (BC) management is the identification of biomarkers for disease aggressiveness. Urine is a "gold mine" for biomarker discovery, nevertheless, with multiple proteins being in low amounts, urine proteomics becomes challenging. In the present study we applied a fractionation strategy of urinary proteins based on the use of immobilized metal affinity chromatography for the discovery of biomarkers for aggressive BC. Urine samples from patients with non invasive (two pools) and invasive (two pools) BC were subjected to immobilized metal affinity chromatography fractionation and eluted proteins analyzed by 1D-SDS-PAGE, band excision and liquid chromatography tandem MS. Among the identified proteins, multiple corresponded to proteins with affinity for metals and/or reported to be phosphorylated and included proteins with demonstrated association with BC such as MMP9, fibrinogen forms, and clusterin. In agreement to the immobilized metal affinity chromatography results, aminopeptidase N, profilin 1, and myeloblastin were further found to be differentially expressed in urine from patients with invasive compared with non invasive BC and benign controls, by Western blot or Elisa analysis, nevertheless exhibiting high interindividual variability. By tissue microarray analysis, profilin 1 was found to have a marked decrease of expression in the epithelial cells of the invasive (T2+) versus high risk non invasive (T1G3) tumors with occasional expression in stroma; importantly, this pattern strongly correlated with poor prognosis and increased mortality. The functional relevance of profilin 1 was investigated in the T24 BC cells where blockage of the protein by the use of antibodies resulted in decreased cell motility with concomitant decrease in actin polymerization. Collectively, our study involves the application of a fractionation method of urinary proteins and as one main result of this analysis reveals the association of profilin 1 with BC paving the way for its further investigation in BC stratification. Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.009449, 1-15, 2012.