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[iTRAQ] 肾透明细胞癌-iTRAQ-biomarker-new therapies

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发表于 2015-12-28 14:49:15 | 显示全部楼层 |阅读模式

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Oncotarget. 2014 Jan 30;5(2):506-18.
Quantitative proteomic analysis reveals potential diagnostic markers and pathways involved in pathogenesis ofrenal cell carcinoma.IF=
14年6.359
13年6.627
12年6.636

研究关键词 肾透明细胞癌,iTRAQ,biomarker,治疗靶点

研究概要
    目前肾透明细胞癌(ccRCC)的诊断还没有准确的血清学标记。本研究通过定量蛋白质组学的方法,用iTraq进行同位素标记,并进行LC-MS/MS分析。共199个患者被纳入研究,iTRAQ检测发现在ccRCC样本中有55个蛋白的表达量与正常肾组织有显著差异。其中54个蛋白曾经被报导在癌变过程中发挥作用,有39个蛋白是分泌蛋白。失调的蛋白中,alpha-enolase(ENO1), L-lactate dehydrogenase A chain (LDHA), heat shock protein beta-1 (HSPB1/Hsp27), 10 kDa heat shock protein, mitochondrial (HSPE1)的差异在另外两组独立病人中应用WesternblotIHC得到验证。用PCR进行的信号通路分析验证表明,葡萄糖代谢在ccRCC病人中相比正常肾组织有所改变。作为备选的诊断标记,我们检查了Hsp27在血清和尿液中的变化。在ccRCC病人中,Hsp27在血清和尿液中显著升高,Hsp27的升高与肿瘤的等级有关。这些数据显示揭示了ccRCC癌变过程的机制,并发现了潜在ccRCC诊断标记Hsp27。当然作为诊断标记还需要大规模样本的验证。蛋白谱数据也为发现新的治疗提供的重要线索。


以下为技术路线:

技术路线图.png

Abstract
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 proteomicsanalysis 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.
PMID: 24504108




免费全文链接:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3964225/




Oncotarget.2014.IF6.636.S2.pdf

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Oncotarget.2014.IF6.636.pdf

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