Pilot and feasibility study: prospective proteomic profiling of mammary epithelial cells from high-risk women provides evidence of activation of pro-survival pathways
Pilot and feasibility study: prospective proteomic profiling of mammary epithelial cells from high-risk women provides evidence of activation of pro-survival pathways
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DOI：
10.1007/s10549-011-1609-9
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发表时间：
2012-04-01
影响因子：
3.8
通讯作者：
Seewaldt, Victoria L.
中科院分区：
文献类型：
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作者：
Ibarra-Drendall, Catherine;Troch, Michelle M.;Seewaldt, Victoria L.
正常的乳腺稳态需要蛋白质信号网络的协调调节。然而，我们有一点前瞻性的信息，蛋白质信号的激活是否发生在癌前乳腺上皮细胞，代表细胞与乳腺癌高危妇女的细胞学异常。这些信息对于理解解除调节的信号通路在乳腺癌发生中的作用以及将来制定针对性的乳腺癌预防和/或治疗策略至关重要。在这项试点和可行性研究中，我们研究了52磷酸化，总，和切割蛋白质的表达在31微解剖随机乳晕细针穿刺（RPFNA）样本的高通量反相蛋白质微阵列。无监督分层聚类分析表明存在代表以下信号通路的四个蛋白质簇：（1）受体酪氨酸激酶/Akt/哺乳动物雷帕霉素靶蛋白（RTK/Akt/mTOR），（2）RTK/Akt/细胞外信号调节激酶（RTK/Akt/ERK），（3）线粒体凋亡，和（4）不确定。簇1至3包含中度至高度表达的蛋白质，而簇4包含在大多数RPFNA样品中低表达的蛋白质。我们的探索性研究表明，线粒体细胞凋亡途径的相互关联的成分在从高危女性获得的所有乳腺上皮细胞中高度表达。特别地，抗凋亡Bcl-xL和促凋亡Bad的表达水平在非典型和非典型样品中均呈正相关（未校正的P < 0.0001），表明在乳腺癌发生的早期步骤期间细胞增殖的促凋亡和抗凋亡调节之间存在微妙的平衡。我们的可行性研究表明，沿着RTK/Akt通路的关键蛋白的激活可能会使这种平衡对细胞存活产生影响。综上所述，我们的研究结果表明，在从高危女性获得的有限RPFNA样本中映射蛋白质组信号网络的可行性，以及在未来的蛋白质组学研究中开发合理的药物靶点或乳腺癌预防策略的前景。
Normal mammary gland homeostasis requires the coordinated regulation of protein signaling networks. However, we have little prospective information on whether activation of protein signaling occurs in premalignant mammary epithelial cells, as represented by cells with cytological atypia from women who are at high risk for breast cancer. This information is critical for understanding the role of deregulated signaling pathways in the initiation of breast cancer and for developing targeted prevention and/or treatment strategies for breast cancer in the future. In this pilot and feasibility study, we examined the expression of 52 phosphorylated, total, and cleaved proteins in 31 microdissected Random Periareolar Fine Needle Aspiration (RPFNA) samples by high-throughput Reverse Phase Protein Microarray. Unsupervised hierarchical clustering analysis indicated the presence of four clusters of proteins that represent the following signaling pathways: (1) receptor tyrosine kinase/Akt/mammalian target of rapamycin (RTK/Akt/mTOR), (2) RTK/Akt/extracellular signal-regulated kinase (RTK/Akt/ERK), (3) mitochondrial apoptosis, and (4) indeterminate. Clusters 1 through 3 comprised moderately to highly expressed proteins, while Cluster 4 comprised proteins that are lowly expressed in a majority of RPFNA samples. Our exploratory study showed that the interlinked components of mitochondrial apoptosis pathway are highly expressed in all mammary epithelial cells obtained from high-risk women. In particular, the expression levels of anti-apoptotic Bcl-xL and pro-apoptotic Bad are positively correlated in both non-atypical and atypical samples (unadjusted P < 0.0001), suggesting a delicate balance between the pro-apoptotic and anti-apoptotic regulation of cell proliferation during the early steps of mammary carcinogenesis. Our feasibility study suggests that the activation of key proteins along the RTK/Akt pathway may tip this balance to cell survival. Taken together, our results demonstrate the feasibility of mapping proteomic signaling networks in limited RPFNA samples obtained from high-risk women and the promise of developing rational drug targets or preventative strategies for breast cancer in future proteomic studies with a larger cohort of high-risk women.