Gene expression signatures and biomarkers of noninvasive and invasive breast cancer cells: comprehensive profiles by representational difference analysis, microarrays and proteomics
Gene expression signatures and biomarkers of noninvasive and invasive breast cancer cells: comprehensive profiles by representational difference analysis, microarrays and proteomics
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DOI：
10.1038/sj.onc.1209265
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发表时间：
2006-04-13
期刊：
影响因子：
8
通讯作者：
Kandpal, RP
中科院分区：
文献类型：
--
作者：
Nagaraja, GM;Othman, M;Kandpal, RP
我们已经表征了对应于正常乳腺（MCF 10A）、非侵袭性乳腺癌（MCF 7）和侵袭性乳腺癌（MDA-MB-231）的细胞系的全面转录和蛋白质组学谱。首先通过MCF 7和MDA-MB-231细胞之间cDNA的代表性差异分析（RDA）的修改方案分析转录物谱。RDA鉴定的大多数基因与微阵列结果几乎完全一致，并且还鉴定了一些差异表达的基因，如赖氨酰氧化酶、铜转运蛋白ATP 7A、EphB 6、RUNX 2和RUNX 2的变体。通过微阵列分析鉴定的改变的转录物涉及细胞-细胞或细胞-基质相互作用、Rho信号传导、钙稳态和铜结合/敏感活性。包括GPCR 11、钙粘蛋白11、膜联蛋白A1、波形蛋白、乳酸脱氢酶B（在MDA-MB-231中上调）和GREB 1、S100 A8、淀粉样β前体蛋白、密蛋白3和钙粘蛋白1（在MDA-MB-231中下调）在内的一组9个基因足以区分MDA-MB-231和MCF 7细胞。下调一组转录的蛋白质参与细胞-细胞相互作用表明这些转录作为潜在的标记物的侵袭性，可以通过甲基化特异性PCR检测。蛋白质组学谱表明与转录谱相比，较少蛋白质的丰度改变。反义敲除选定的转录导致细胞增殖的抑制，伴随着改变蛋白质组学谱。反义转染子的蛋白质组学特征表明肽基脯氨酰异构酶、Raf激酶抑制剂和80 kDa蛋白激酶C底物参与介导细胞增殖的抑制。
We have characterized comprehensive transcript and proteomic profiles of cell lines corresponding to normal breast (MCF10A), noninvasive breast cancer (MCF7) and invasive breast cancer (MDA-MB-231). The transcript profiles were first analysed by a modified protocol for representational difference analysis (RDA) of cDNAs between MCF7 and MDA-MB-231 cells. The majority of genes identified by RDA showed nearly complete concordance with microarray results, and also led to the identification of some differentially expressed genes such as lysyl oxidase, copper transporter ATP7A, EphB6, RUNX2 and a variant of RUNX2. The altered transcripts identified by microarray analysis were involved in cell-cell or cell-matrix interaction, Rho signaling, calcium homeostasis and copper-binding/sensitive activities. A set of nine genes that included GPCR11, cadherin 11, annexin A1, vimentin, lactate dehydrogenase B (upregulated in MDA-MB-231) and GREB1, S100A8, amyloid beta precursor protein, claudin 3 and cadherin 1 (downregulated in MDA-MB-231) were sufficient to distinguish MDA-MB-231 from MCF7 cells. The downregulation of a set of transcripts for proteins involved in cell-cell interaction indicated these transcripts as potential markers for invasiveness that can be detected by methylation-specific PCR. The proteomic profiles indicated altered abundance of fewer proteins as compared to transcript profiles. Antisense knockdown of selected transcripts led to inhibition of cell proliferation that was accompanied by altered proteomic profiles. The proteomic profiles of antisense transfectants suggest the involvement of peptidylprolyl isomerase, Raf kinase inhibitor and 80 kDa protein kinase C substrate in mediating the inhibition of cell proliferation.