PMID- 23762360 OWN - NLM STAT- MEDLINE DCOM- 20140211 LR - 20211021 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 8 IP - 6 DP - 2013 TI - Synergism between Hedgehog-GLI and EGFR signaling in Hedgehog-responsive human medulloblastoma cells induces downregulation of canonical Hedgehog-target genes and stabilized expression of GLI1. PG - e65403 LID - 10.1371/journal.pone.0065403 [doi] LID - e65403 AB - Aberrant activation of Hedgehog (HH) signaling has been identified as a key etiologic factor in many human malignancies. Signal strength, target gene specificity, and oncogenic activity of HH signaling depend profoundly on interactions with other pathways, such as epidermal growth factor receptor-mediated signaling, which has been shown to cooperate with HH/GLI in basal cell carcinoma and pancreatic cancer. Our experimental data demonstrated that the Daoy human medulloblastoma cell line possesses a fully inducible endogenous HH pathway. Treatment of Daoy cells with Sonic HH or Smoothened agonist induced expression of GLI1 protein and simultaneously prevented the processing of GLI3 to its repressor form. To study interactions between HH- and EGF-induced signaling in greater detail, time-resolved measurements were carried out and analyzed at the transcriptomic and proteomic levels. The Daoy cells responded to the HH/EGF co-treatment by downregulating GLI1, PTCH, and HHIP at the transcript level; this was also observed when Amphiregulin (AREG) was used instead of EGF. We identified a novel crosstalk mechanism whereby EGFR signaling silences proteins acting as negative regulators of HH signaling, as AKT- and ERK-signaling independent process. EGFR/HH signaling maintained high GLI1 protein levels which contrasted the GLI1 downregulation on the transcript level. Conversely, a high-level synergism was also observed, due to a strong and significant upregulation of numerous canonical EGF-targets with putative tumor-promoting properties such as MMP7, VEGFA, and IL-8. In conclusion, synergistic effects between EGFR and HH signaling can selectively induce a switch from a canonical HH/GLI profile to a modulated specific target gene profile. This suggests that there are more wide-spread, yet context-dependent interactions, between HH/GLI and growth factor receptor signaling in human malignancies. FAU - Gotschel, Frank AU - Gotschel F AD - Division of Molecular Genome Analysis, German Cancer Research Center-DKFZ, Heidelberg, Germany. FAU - Berg, Daniela AU - Berg D FAU - Gruber, Wolfgang AU - Gruber W FAU - Bender, Christian AU - Bender C FAU - Eberl, Markus AU - Eberl M FAU - Friedel, Myriam AU - Friedel M FAU - Sonntag, Johanna AU - Sonntag J FAU - Rungeler, Elena AU - Rungeler E FAU - Hache, Hendrik AU - Hache H FAU - Wierling, Christoph AU - Wierling C FAU - Nietfeld, Wilfried AU - Nietfeld W FAU - Lehrach, Hans AU - Lehrach H FAU - Frischauf, Annemarie AU - Frischauf A FAU - Schwartz-Albiez, Reinhard AU - Schwartz-Albiez R FAU - Aberger, Fritz AU - Aberger F FAU - Korf, Ulrike AU - Korf U LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20130610 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Biomarkers, Tumor) RN - 0 (Carrier Proteins) RN - 0 (Cyclohexylamines) RN - 0 (Enzyme Inhibitors) RN - 0 (GLI1 protein, human) RN - 0 (HHIP protein, human) RN - 0 (Hedgehog Proteins) RN - 0 (Interleukin-8) RN - 0 (Membrane Glycoproteins) RN - 0 (PTCH1 protein, human) RN - 0 (Patched Receptors) RN - 0 (Patched-1 Receptor) RN - 0 (RNA, Messenger) RN - 0 (Receptors, Cell Surface) RN - 0 (SAG compound) RN - 0 (SHH protein, human) RN - 0 (Thiophenes) RN - 0 (Transcription Factors) RN - 0 (VEGFA protein, human) RN - 0 (Vascular Endothelial Growth Factor A) RN - 0 (Veratrum Alkaloids) RN - 0 (Zinc Finger Protein GLI1) RN - EC 2.7.10.1 (EGFR protein, human) RN - EC 2.7.10.1 (ErbB Receptors) RN - EC 3.4.24.23 (MMP7 protein, human) RN - EC 3.4.24.23 (Matrix Metalloproteinase 7) RN - ZH658AJ192 (cyclopamine) SB - IM MH - Biomarkers, Tumor/genetics/metabolism MH - Blotting, Western MH - Carrier Proteins/genetics/metabolism MH - Cell Proliferation MH - Cells, Cultured MH - Cerebellar Neoplasms/drug therapy/*metabolism/pathology MH - Cyclohexylamines/pharmacology MH - Dermis/cytology/drug effects/metabolism MH - Drug Synergism MH - Enzyme Inhibitors/pharmacology MH - Enzyme-Linked Immunosorbent Assay MH - ErbB Receptors/genetics/*metabolism MH - Fibroblasts/cytology/drug effects/metabolism MH - Gene Expression Profiling MH - HEK293 Cells MH - Hedgehog Proteins/agonists/genetics/*metabolism MH - Humans MH - Interleukin-8/genetics/metabolism MH - Matrix Metalloproteinase 7/genetics/metabolism MH - Medulloblastoma/drug therapy/*metabolism/pathology MH - Membrane Glycoproteins/genetics/metabolism MH - Oligonucleotide Array Sequence Analysis MH - Patched Receptors MH - Patched-1 Receptor MH - Phosphorylation/drug effects MH - Protein Array Analysis MH - RNA, Messenger/genetics MH - Real-Time Polymerase Chain Reaction MH - Receptors, Cell Surface/genetics/metabolism MH - Reverse Transcriptase Polymerase Chain Reaction MH - Thiophenes/pharmacology MH - Transcription Factors/genetics/*metabolism MH - Vascular Endothelial Growth Factor A/genetics/metabolism MH - Veratrum Alkaloids/pharmacology MH - Zinc Finger Protein GLI1 PMC - PMC3677915 COIS- Competing Interests: The authors have declared that no competing interests exist. EDAT- 2013/06/14 06:00 MHDA- 2014/02/12 06:00 PMCR- 2013/06/10 CRDT- 2013/06/14 06:00 PHST- 2013/01/24 00:00 [received] PHST- 2013/04/24 00:00 [accepted] PHST- 2013/06/14 06:00 [entrez] PHST- 2013/06/14 06:00 [pubmed] PHST- 2014/02/12 06:00 [medline] PHST- 2013/06/10 00:00 [pmc-release] AID - PONE-D-13-04483 [pii] AID - 10.1371/journal.pone.0065403 [doi] PST - epublish SO - PLoS One. 2013 Jun 10;8(6):e65403. doi: 10.1371/journal.pone.0065403. Print 2013.