RNA-Seq of human breast ductal carcinoma in situ models reveals aldehyde dehydrogenase isoform 5A1 as a novel potential target.
RNA-Seq of human breast ductal carcinoma in situ models reveals aldehyde dehydrogenase isoform 5A1 as a novel potential target.
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DOI：
10.1371/journal.pone.0050249
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Mattingly RR
中科院分区：
文献类型：
--
作者：
Kaur H;Mao S;Li Q;Sameni M;Krawetz SA;Sloane BF;Mattingly RR
乳腺导管原位癌（DCIS）被发现在大量的妇女由于广泛使用的乳房X线摄影。为了增加对DCIS的了解，我们确定了在具有重构基底膜（rBM）的三维（3D）覆盖培养中，与非致瘤性乳腺上皮细胞的MCF 10A模型相比，在三种DCIS模型（MCF 10.DCIS、SUM 102和SUM 225）中常见的表达变化。使用Illumina基因组分析仪GAIIx对提取的mRNA进行76个循环的深度测序（RNA-Seq）。RNA-Seq结果分析显示，DCIS模型中有295个一致差异表达的转录本。这些差异表达的基因编码的蛋白质与许多信号传导途径相关，如整联蛋白、成纤维细胞生长因子和TGFβ信号传导，显示与细胞间信号传导、细胞间粘附和细胞增殖相关，并具有明显的细胞外和质膜区室定位偏好。通过选择的差异表达基因的定量实时PCR验证RNA-Seq数据。乙醛脱氢酶5A 1（ALDH 5A 1）是一种参与线粒体谷氨酸代谢的酶，在所有三种DCIS模型中在mRNA和蛋白质水平均过表达。已知双硫仑和丙戊酸可抑制ALDH 5A 1，并且可安全地长期用于人类其他疾病。这两种药物均显著抑制DCIS 3D rBM覆盖模型的净增殖，但对MCF 10A 3D rBM覆盖模型的影响极小。这些结果表明，ALDH 5A 1可能在DCIS中发挥重要作用，并可能成为一种新的分子治疗靶点。
Breast ductal carcinoma in situ (DCIS) is being found in great numbers of women due to the widespread use of mammography. To increase knowledge of DCIS, we determined the expression changes that are common among three DCIS models (MCF10.DCIS, SUM102 and SUM225) compared to the MCF10A model of non-tumorigenic mammary epithelial cells in three dimensional (3D) overlay culture with reconstituted basement membrane (rBM). Extracted mRNA was subjected to 76 cycles of deep sequencing (RNA-Seq) using Illumina Genome Analyzer GAIIx. Analysis of RNA-Seq results showed 295 consistently differentially expressed transcripts in the DCIS models. These differentially expressed genes encode proteins that are associated with a number of signaling pathways such as integrin, fibroblast growth factor and TGFβ signaling, show association with cell-cell signaling, cell-cell adhesion and cell proliferation, and have a notable bias toward localization in the extracellular and plasma membrane compartments. RNA-Seq data was validated by quantitative real-time PCR of selected differentially expressed genes. Aldehyde dehydrogenase 5A1 (ALDH5A1) which is an enzyme that is involved in mitochondrial glutamate metabolism, was over-expressed in all three DCIS models at both the mRNA and protein levels. Disulfiram and valproic acid are known to inhibit ALDH5A1 and are safe for chronic use in humans for other disorders. Both of these drugs significantly inhibited net proliferation of the DCIS 3D rBM overlay models, but had minimal effect on MCF10A 3D rBM overlay models. These results suggest that ALDH5A1 may play an important role in DCIS and potentially serve as a novel molecular therapeutic target.
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影响因子：
4.8
作者：
Debnath, J;Muthuswamy, SK;Brugge, JS
通讯作者：
Brugge, JS
影响因子：
5.6
作者：
Eyal, S;Yagen, B;Bialer, M
通讯作者：
Bialer, M
影响因子：
3.4
作者：
Han, K.;Nofech-Mozes, S.;Rakovitch, E.
通讯作者：
Rakovitch, E.
DOI：
10.1186/bcr2923
发表时间：
2011
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Griffith OL;Gray JW
通讯作者：
Gray JW
DOI：
10.1186/bcr1613
发表时间：
2006
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Hannemann J;Velds A;Halfwerk JB;Kreike B;Peterse JL;van de Vijver MJ
通讯作者：
van de Vijver MJ