Quantitative proteomic analysis reveals potential diagnostic markers and pathways involved in pathogenesis of renal cell carcinoma.
Quantitative proteomic analysis reveals potential diagnostic markers and pathways involved in pathogenesis of renal cell carcinoma.
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DOI：
10.18632/oncotarget.1529
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发表时间：
2014-01-30
期刊：
影响因子：
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通讯作者：
Yousef GM
中科院分区：
文献类型：
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作者：
White NM;Masui O;Desouza LV;Krakovska O;Metias S;Romaschin AD;Honey RJ;Stewart R;Pace K;Lee J;Jewett MA;Bjarnason GA;Siu KW;Yousef GM
目前尚无准确诊断肾透明细胞癌(CcRCC)的血清标志物。肾切除术的诊断和决定依赖于影像，但影像并不总是准确的。迫切需要非侵入性的诊断生物标志物。在这项研究中，我们对总共199例患者进行了定量蛋白质组学分析，其中包括30对配对的正常肾和肾细胞癌，使用等压标记进行相对和绝对定量(ITRAQ)标记和LC-MS/MS分析以确定差异表达的蛋白质。我们发现，与正常肾组织相比，ccRCC中有55个蛋白表达明显异常。此前有54个蛋白被报道在癌症发生中起作用，39个是分泌型蛋白。免疫印迹和免疫组织化学检测证实两组患者存在α-烯醇化酶(ENO1)、L乳酸脱氢酶A链(LDHA)、热休克蛋白β1(HSPB1/HSP27)和10 kDa热休克蛋白线粒体(HSPE1)的异常表达。经聚合酶链式反应验证的通路分析表明，与正常肾组织相比，肾小管细胞癌的葡萄糖代谢发生了改变。此外，我们还检测了Hsp27在血清和尿液中作为生物标志物的应用。在肾细胞癌患者中，尿和血清Hsp27水平升高，且血清Hsp27水平升高与高级别(3~4级)肿瘤相关。这些数据共同确定了ccRCC潜在的诊断生物标志物，并为阐明调控失调并有助于ccRCC发病机制的分子机制提供了新的线索。Hsp27是一种有前景的肾细胞癌诊断标记物，但仍需进一步的大规模研究。此外，分子图谱可能有助于为新疗法的发现铺平道路。
There are no serum biomarkers for the accurate diagnosis of clear cell renal cell carcinoma (ccRCC). Diagnosis and decision of nephrectomy rely on imaging which is not always accurate. Non-invasive diagnostic biomarkers are urgently required. In this study, we preformed quantitative proteomics analysis on a total of 199 patients including 30 matched pairs of normal kidney and ccRCC using isobaric tags for relative and absolute quantitation (iTRAQ) labeling and LC-MS/MS analysis to identify differentially expressed proteins. We found 55 proteins significantly dysregulated in ccRCC compared to normal kidney tissue. 54 were previously reported to play a role in carcinogenesis, and 39 are secreted proteins. Dysregulation of alpha-enolase (ENO1), L-lactate dehydrogenase A chain (LDHA), heat shock protein beta-1 (HSPB1/Hsp27), and 10 kDa heat shock protein, mitochondrial (HSPE1) was confirmed in two independent sets of patients by western blot and immunohistochemistry. Pathway analysis, validated by PCR, showed glucose metabolism is altered in ccRCC compared to normal kidney tissue. In addition, we examined the utility of Hsp27 as biomarker in serum and urine. In ccRCC patients, Hsp27 was elevated in the urine and serum and high serum Hsp27 was associated with high grade (Grade 3–4) tumors. These data together identify potential diagnostic biomarkers for ccRCC and shed new light on the molecular mechanisms that are dysregulated and contribute to the pathogenesis of ccRCC. Hsp27 is a promising diagnostic marker for ccRCC although further large-scale studies are required. Also, molecular profiling may help pave the road to the discovery of new therapies.