Phosphoglucose isomerase/autocrine motility factor mediates epithelial-mesenchymal transition regulated by miR-200 in breast cancer cells.
Phosphoglucose isomerase/autocrine motility factor mediates epithelial-mesenchymal transition regulated by miR-200 in breast cancer cells.
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DOI：
10.1158/0008-5472.can-10-0965
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发表时间：
2011-05-01
期刊：
影响因子：
11.2
通讯作者：
Raz A
中科院分区：
文献类型：
--
作者：
Ahmad A;Aboukameel A;Kong D;Wang Z;Sethi S;Chen W;Sarkar FH;Raz A
磷酸葡萄糖异构酶/自分泌运动因子（PGI/AMF）在糖酵解和肿瘤发生中起重要作用，与肿瘤细胞的侵袭和转移有关。我们之前已经显示了它在乳腺癌细胞中诱导上皮向间充质转化（EMT）中的作用，这导致了侵袭性增加;然而，PGI/AMF调节EMT的分子机制尚不清楚。我们首次发现PGI/AMF过表达导致NF-κB的DNA结合活性增加，进而导致ZEB 1/ZEB 2表达增加。已知microRNA-200 s（miR-200 s; miR-200 a，miR-200 b和miR-200 c）负调控ZEB 1/ZEB 2的表达，我们发现miR-200 s的表达在PGI/AMF过表达的MCF-10A细胞以及高度侵袭性的MDA-MB-231细胞中丢失，这与ZEB 1/ZEB 2的表达增加一致。此外，MDA-MB-231细胞中PGI/AMF表达的沉默导致miR-200的过度表达，这与EMT表型即间质向上皮转化（MET）的逆转相关，并且这些发现与上皮细胞相对表达的改变一致。（E-钙粘蛋白）和间充质（波形蛋白，ZEB 1，ZEB 2）标记物，以及通过克隆形成、运动性和侵袭性测定判断的侵袭性降低。此外，miR-200的再表达或PGI/AMF的沉默抑制了体内MDA-MB-231细胞的肺转移，并且体内抗miR-200治疗导致转移增加。总之，这些结果表明miR-200在PGI/AMF诱导的EMT中的作用，因此上调miR-200的方法可能是治疗高度侵袭性乳腺癌的新治疗策略。
Phosphoglucose isomerase/autocrine motility factor (PGI/AMF) plays important role in glycolysis and gluconeogenesis, and is associated with invasion and metastasis of cancer cells. We have previously shown its role in the induction of Epithelial-to-Mesenchymal transition (EMT) in breast cancer cells, which led to increased aggressiveness; however, the molecular mechanism by which PGI/AMF regulates EMT is not known. Here we show, for the first time, that PGI/AMF over-expression led to an increase in the DNA-binding activity of NF-κB, which, in turn, led to increased expression of ZEB1/ZEB2. The microRNA-200s (miR-200s; miR-200a, miR-200b and miR-200c) are known to negatively regulate the expression of ZEB1/ZEB2, and we found that the expression of miR-200s was lost in PGI/AMF over-expressing MCF-10A cells as well as in highly invasive MDA-MB-231 cells, which was consistent with increased expression of ZEB1/ZEB2. Moreover, silencing of PGI/AMF expression in MDA-MB-231 cells led to over-expression of miR-200s, which was associated with reversal of EMT phenotype i.e. Mesenchymal-to-Epithelial Transition (MET), and these findings were consistent with alterations in the relative expression of epithelial (E-cadherin) and mesenchymal (vimentin, ZEB1, ZEB2) markers, and decreased aggressiveness as judged by clonogenic, motility and invasion assays. Moreover, either re-expression of miR-200 or silencing of PGI/AMF suppressed pulmonary metastases of MDA-MB-231 cells in vivo, and anti-miR-200 treatment in vivo resulted in increased metastases. Collectively, these results suggest a role of miR-200s in PGI/AMF induced EMT, and thus approaches for up-regulation of miR-200s could be a novel therapeutic strategy for the treatment of highly invasive breast cancer.