NF-κB represses E-cadherin expression and enhances epithelial to mesenchymal transition of mammary epithelial cells:: potential involvement of ZEB-1 and ZEB-2
NF-κB represses E-cadherin expression and enhances epithelial to mesenchymal transition of mammary epithelial cells:: potential involvement of ZEB-1 and ZEB-2
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DOI：
10.1038/sj.onc.1209808
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发表时间：
2007-02-01
期刊：
影响因子：
8
通讯作者：
Nakshatri, H.
中科院分区：
文献类型：
--
作者：
Chua, H. L.;Bhat-Nakshatri, P.;Nakshatri, H.
转录因子核因子κ B（NF-κ B）在乳腺癌的癌细胞和基质细胞中都具有组成性活性;然而，活化的NF-κ B在癌症进展中的确切作用尚不清楚。使用亲本MCF 10A细胞和一种表达肌上皮标志物p63的变体，该变体稳定过表达NF-κ B的组成型活性p65亚基（MCF 10A/p65），我们表明NF-κ B抑制上皮特异性基因E-钙粘蛋白和桥粒斑蛋白的表达，并诱导间充质特异性基因波形蛋白的表达。P65还抑制p63和假定的乳腺上皮祖细胞标记物细胞角蛋白5/6的表达。MCF 10A/p65细胞与经历上皮向间充质转化（EMT）的细胞表型相似。MCF 10A/p65细胞在三维Matrigel中不能形成特征性的腺泡。对亲本和MCF 10A/p65细胞中先前显示参与EMT的基因的分析显示，与亲本细胞相比，MCF 10A/p65细胞中ZEB-1和ZEB-2的表达升高。在瞬时转染试验中，p65增加ZEB-1启动子活性。此外，过表达ZEB-1的MCF-10A细胞显示E-钙粘蛋白和p63表达减少，并显示EMT表型。针对ZEB-1或ZEB-2的siRNA减少了存活的MCF 10A/p65细胞的数量，但不减少亲本细胞的数量，表明MCF 10A/p65细胞依赖于ZEB-1和ZEB-2的细胞周期进展或存活。长期暴露于肿瘤坏死因子α（TNF α）（一种有效的NF-κ B诱导剂）的MCF 10 A细胞也表现出EMT样表型和ZEB-1/ZEB-2诱导，这两者在TNF α撤除后都被逆转。
The transcription factor nuclear factor kappa B (NF-kappa B) is constitutively active in both cancer cells and stromal cells of breast cancer; however, the precise role of activated NF-kappa B in cancer progression is not known. Using parental MCF10A cells and a variant that expresses the myoepithelial marker p63 stably overexpressing the constitutively active p65 subunit of NF-kappa B (MCF10A/p65), we show that NF-kappa B suppresses the expression of epithelial specific genes E-cadherin and desmoplakin and induces the expression of the mesenchymal specific gene vimentin. P65 also suppressed the expression of p63 and the putative breast epithelial progenitor marker cytokeratin 5/6. MCF10A/p65 cells were phenotypically similar to cells undergoing epithelial to mesenchymal transition (EMT). MCF10A/p65 cells failed to form characteristic acini in three-dimensional Matrigel. Analysis of parental and MCF10A/p65 cells for genes previously shown to be involved in EMT revealed elevated expression of ZEB-1 and ZEB-2 in MCF10A/p65 cells compared to parental cells. In transient transfection assays, p65 increased ZEB-1 promoter activity. Furthermore, MCF10A cells overexpressing ZEB-1 showed reduced E-cadherin and p63 expression and displayed an EMT phenotype. The siRNA against ZEB-1 or ZEB-2 reduced the number of viable MCF10A/p65 but not parental cells, suggesting the dependence of MCF10A/p65 cells to ZEB-1 and ZEB-2 for cell cycle progression or survival. MCF10A cells chronically exposed to tumor necrosis factor alpha (TNF alpha), a potent NF-kappa B inducer, also exhibited the EMT-like phenotype and ZEB-1/ZEB-2 induction, both of which were reversed following TNFa withdrawal.