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nature portfolio privacy policy advertising nature 449 nature 392 nature 451 nature author information authors invasion kits 0/ reprints notch/hif/maml1 immuno-complex initiative event resulting icn3-induced notch signaling regulates notch signaling human ras-transformed cells warm serum-free media ligand-induced notch signaling inhibits hras1-induced oncogenesis hypoxia inducible factor-1α hrp-conjugated secondary antibodies author correspondence dana-farber cancer institute notch1-induced mammary tumorigenesis ligand-mediated notch activation rbp-jκ-binding sites rbp-jκ binding sites real-time pcr analysis published pathway figures zinc-finger proteins snail mouse mammary epithelium cell–cell adhesion molecule hypoxia-mediated notch signaling pre-existing notch signaling real-time pcr experiments induces mammary tumors notch-dependent mechanism hif-1α transcription complex hif-2α potentiated icn1 mouse mammary tumor scratch wound-healing assay low oxygen content antibodies pcdna3/hif-1α full size image notch intracellular domain breast cancer development hypoxia inducible factor permissions notch signaling induces notch1 intracellular domain human mammary tumourigenesis

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• Peinado H, Olmeda D, Cano A (2007) Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype?

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         description:Epithelial-to-mesenchymal transition (EMT) is associated with decreased adhesion and acquisition of metastatic potential of breast cancer cells. Epithelial-to-mesenchymal transition is mediated, in part, by two transcription repressors, Snail and Slug, that are known to be targets of the Notch signaling pathway, and JAGGED1-induced Notch activation increases EMT. However, the events that lead to increased Notch activity during EMT of breast cancer cells are unknown. The accumulation of hypoxia inducible factors (HIFs) under hypoxia was detected by western blot analysis, and their effects on Notch signaling were measured by an in vitro Notch reporter assay. The expression of Notch target genes under hypoxia was tested by real-time PCR. The knockdown of HIF-1α was mediated by retroviral delivery of shRNA. The expression of Slug and Snail under hypoxia was measured by real-time PCR. Breast cancer cell migration and invasion under hypoxia were tested with cell migration and invasion kits. Hypoxia increased the expression of Notch target genes such as HES1 and HEY1 in breast cancer cells, as was expression of Notch receptors and ligands. The mechanism is likely to involve the accumulation of HIF-1α and HIF-2α in these cells by hypoxia, which synergised with the Notch co-activator MAML1 in potentiating Notch activity. Hypoxia inducible factor-1α was found to bind to HES1 promoter under hypoxia. Knockdown of HIF-1α with shRNA inhibited both HES1 and HEY1 expression under hypoxia. Hypoxia increased the expression of Slug and Snail, and decreased the expression of E-cadherin, hallmarks of EMT. Notch pathway inhibition abrogated the hypoxia-mediated increase in Slug and Snail expression, as well as decreased breast cancer cell migration and invasion. Hypoxia-mediated Notch signaling may have an important role in the initiation of EMT and subsequent potential for breast cancer metastasis.
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      headline:Hypoxia potentiates Notch signaling in breast cancer leading to decreased E-cadherin expression and increased cell migration and invasion
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