Eight-Plex iTRAQ Analysis of Variant Metastatic Human Prostate Cancer Cells Identifies Candidate Biomarkers of Progression: An Exploratory Study
Eight-Plex iTRAQ Analysis of Variant Metastatic Human Prostate Cancer Cells Identifies Candidate Biomarkers of Progression: An Exploratory Study
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DOI：
10.1002/pros.21167
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发表时间：
2010-09-01
期刊：
影响因子：
2.8
通讯作者：
Rehman, Ishtiaq
中科院分区：
文献类型：
--
作者：
Glen, Adam;Evans, Caroline A.;Rehman, Ishtiaq
背景。由于前列腺癌生物学行为的异质性，迫切需要能够可靠区分惰性和侵袭性疾病的生物标志物来指导治疗选择。我们使用8-plex等压标签进行相对和绝对定量（iTRAQ），以分析具有不同生长和转移潜力的两组不同的等基因前列腺癌细胞的蛋白质组，以确定与进展相关的新生物标志物。LNCaP、LNCaP- pro5和LNCaP- ln3细胞组为雄激素应答型前列腺癌模型，PC-3、PC-3M和PC-3M- ln4细胞组为雄激素不敏感型前列腺癌模型。在鉴定和定量的245个独特蛋白中（>= 95%置信度；>= 2个肽/蛋白），17个在至少一个变异LNCaP细胞中相对于亲代细胞表现出显著的差异表达（>=+/- 1.5）。同样地，在PC-3小组内的比较鉴定出45种蛋白质，与亲代细胞相比，在至少一种变体PC-3细胞中表现出显著的差异表达。通过Western blotting或免疫细胞化学验证所选候选细胞的差异表达，并通过实时荧光定量PCR （qRT-PCR）检测相应mRNA的表达。前列腺组织微阵列免疫染色发现，与非恶性上皮相比，ERp5在恶性前病变中的免疫表达显著增加（P < 0.0001， Mann Whitney u检验），在高Gleason分级（4-5）和低分级（2-3）癌症中的免疫表达显著增加（P < 0.05）。我们的研究为应用8-plex iTRAQ方法发现前列腺癌进展的临床相关候选生物标志物提供了原理证明。中华医学会医学分会，2011,32(2):1 - 4。(C) 2010 Wiley-Liss, Inc。
BACKGROUND. Due to the heterogeneity in the biological behavior of prostate cancer, biomarkers that can reliably distinguish indolent from aggressive disease are urgently needed to inform treatment choices.METHODS. We employed 8-plex isobaric Tags for Relative and Absolute Quantitation (iTRAQ), to profile the proteomes of two distinct panels of isogenic prostate cancer cells with varying growth and metastatic potentials, in order to identify novel biomarkers associated with progression. The LNCaP, LNCaP-Pro5, and LNCaP-LN3 panel of cells represent a model of androgen-responsive prostate cancer, while the PC-3, PC-3M, and PC-3M-LN4 panel represent a model of androgen-insensitive disease.RESULTS. Of the 245 unique proteins identified and quantified (>= 95% confidence; >= 2 peptides/protein), 17 showed significant differential expression (>=+/- 1.5), in at least one of the variant LNCaP cells relative to parental cells. Similarly, comparisons within the PC-3 panel identified 45 proteins to show significant differential expression in at least one of the variant PC-3 cells compared with parental cells. Differential expression of selected candidates was verified by Western blotting or immunocytochemistry, and corresponding mRNA expression was determined by quantitative real-time PCR (qRT-PCR). Immunostaining of prostate tissue microarrays for ERp5, one of the candidates identified, showed a significant higher immunoexpression in pre-malignant lesions compared with non-malignant epithelium (P < 0.0001, Mann Whitney U-test), and in high Gleason grade (4-5) versus low grade (2-3) cancers (P < 0.05).CONCLUSIONS. Our study provides proof of principle for the application of an 8-plex iTRAQ approach to uncover clinically relevant candidate biomarkers for prostate cancer progression. Prostate 70: 1313-1332, 2010. (C) 2010 Wiley-Liss, Inc.