Identification of Molecular Distinctions Between Normal Breast-Associated Fibroblasts and Breast Cancer-Associated Fibroblasts
Identification of Molecular Distinctions Between Normal Breast-Associated Fibroblasts and Breast Cancer-Associated Fibroblasts
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DOI：
10.1007/s12307-008-0017-0
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发表时间：
2009-12-01
影响因子：
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通讯作者：
Frost, Andra R.
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文献类型：
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作者：
Sadlonova, Andrea;Bowe, Damon B.;Frost, Andra R.
间质成纤维细胞影响乳腺上皮细胞的行为。正常乳腺成纤维细胞(NAF)抑制上皮细胞生长，而乳腺癌成纤维细胞(CAF)抑制生长能力较弱，可促进上皮生长。我们试图找出在NaF和CAF中差异表达的分子，并可能对它们不同的生长调节能力负责。为了确定可溶性分子在成纤维细胞-上皮细胞相互作用中的作用，将NaF与MCF10AT上皮细胞进行3D、Transwell和直接接触共培养。NaF抑制MCF10AT在直接接触和Transwell共培养中的增殖，但在直接共培养中这种抑制作用明显更强，这表明可溶性和接触因素都参与了这一过程。早期传代成纤维细胞培养的基因表达谱鉴定出在NAF和CAF中差异表达的420个基因。8个基因中，FIBULIN 1在NaF中高表达，Dickkopf 1、NeuRegin 1、纤溶酶原激活物抑制物2和组织型纤溶酶原激活物在CAF中高表达。免疫组织化学证实FIBULIN 1在正常乳腺组织中的表达高于癌相关成纤维细胞间质。在乳腺癌中，雌激素受体α阳性的乳腺癌间质中纤维蛋白1的表达较高，而基质中纤维蛋白1的低表达与癌上皮细胞的高增殖有关。总之，NaF和CAF培养的表达谱确定了许多与乳腺癌成纤维细胞-上皮细胞相互作用潜在相关的基因。此外，这些早期传代的成纤维细胞培养可以代表体内间质成纤维细胞的基因表达。
Stromal fibroblasts influence the behavior of breast epithelial cells. Fibroblasts derived from normal breast (NAF) inhibit epithelial growth, whereas fibroblasts from breast carcinomas (CAF) have less growth inhibitory capacity and can promote epithelial growth. We sought to identify molecules that are differentially expressed in NAF versus CAF and potentially responsible for their different growth regulatory abilities. To determine the contribution of soluble molecules to fibroblast-epithelial interactions, NAF were grown in 3D, transwell or direct contact co-cultures with MCF10AT epithelial cells. NAF suppressed proliferation of MCF10AT in both direct contact and transwell co-cultures, but this suppression was significantly greater in direct co-cultures, indicating involvement of both soluble and contact factors. Gene expression profiling of early passage fibroblast cultures identified 420 genes that were differentially expressed in NAF versus CAF. Of the eight genes selected for validation by real-time PCR, FIBULIN 1, was overexpressed in NAF, and DICKKOPF 1, NEUREGULIN 1, PLASMINOGEN ACTIVATOR INHIBITOR 2, and TISSUE PLASMINOGEN ACTIVATOR were overexpressed in CAF. A higher expression of FIBULIN 1 in normal- than cancer-associated fibroblastic stroma was confirmed by immunohistochemistry of breast tissues. Among breast cancers, stromal expression of Fibulin 1 protein was higher in estrogen receptor alpha-positive cancers and low stromal expression of Fibulin 1 correlated with a higher proliferation of cancer epithelial cells. In conclusion, expression profiling of NAF and CAF cultures identified many genes with potential relevance to fibroblast-epithelial interactions in breast cancer. Furthermore, these early passage fibroblast cultures can be representative of gene expression in stromal fibroblasts in vivo.