PMID- 26381405 OWN - NLM STAT- MEDLINE DCOM- 20160422 LR - 20210306 IS - 1083-351X (Electronic) IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 290 IP - 45 DP - 2015 Nov 6 TI - Integrin alpha6beta4 Promotes Autocrine Epidermal Growth Factor Receptor (EGFR) Signaling to Stimulate Migration and Invasion toward Hepatocyte Growth Factor (HGF). PG - 27228-27238 LID - S0021-9258(20)49488-8 [pii] LID - 10.1074/jbc.M115.686873 [doi] AB - Integrin alpha6beta4 is up-regulated in pancreatic adenocarcinomas where it contributes to carcinoma cell invasion by altering the transcriptome. In this study, we found that integrin alpha6beta4 up-regulates several genes in the epidermal growth factor receptor (EGFR) pathway, including amphiregulin (AREG), epiregulin (EREG), and ectodomain cleavage protease MMP1, which is mediated by promoter demethylation and NFAT5. The correlation of these genes with integrin alpha6beta4 was confirmed in The Cancer Genome Atlas Pancreatic Cancer Database. Based on previous observations that integrin alpha6beta4 cooperates with c-Met in pancreatic cancers, we examined the impact of EGFR signaling on hepatocyte growth factor (HGF)-stimulated migration and invasion. We found that AREG and EREG were required for autocrine EGFR signaling, as knocking down either ligand inhibited HGF-mediated migration and invasion. We further determined that HGF induced secretion of AREG, which is dependent on integrin-growth factor signaling pathways, including MAPK, PI3K, and PKC. Moreover, matrix metalloproteinase activity and integrin alpha6beta4 signaling were required for AREG secretion. Blocking EGFR signaling with EGFR-specific antibodies or an EGFR tyrosine kinase inhibitor hindered HGF-stimulated pancreatic carcinoma cell chemotaxis and invasive growth in three-dimensional culture. Finally, we found that EGFR was phosphorylated in response to HGF stimulation that is dependent on EGFR kinase activity; however, c-Met phosphorylation in response to HGF was unaffected by EGFR signaling. Taken together, these data illustrate that integrin alpha6beta4 stimulates invasion by promoting autocrine EGFR signaling through transcriptional up-regulation of key EGFR family members and by facilitating HGF-stimulated EGFR ligand secretion. These signaling events, in turn, promote pancreatic carcinoma migration and invasion. CI - (c) 2015 by The American Society for Biochemistry and Molecular Biology, Inc. FAU - Carpenter, Brittany L AU - Carpenter BL AD - Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0509; Departments of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky 40506-0509. FAU - Chen, Min AU - Chen M AD - Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0509; Departments of Toxicology and Cancer Biology, University of Kentucky, Lexington, Kentucky 40506-0509. FAU - Knifley, Teresa AU - Knifley T AD - Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0509. FAU - Davis, Kelley A AU - Davis KA AD - Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0509. FAU - Harrison, Susan M W AU - Harrison SMW AD - Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0509. FAU - Stewart, Rachel L AU - Stewart RL AD - Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0509; Departments of Pathology and Laboratory Medicine, University of Kentucky, Lexington, Kentucky 40506-0509. FAU - O'Connor, Kathleen L AU - O'Connor KL AD - Markey Cancer Center, University of Kentucky, Lexington, Kentucky 40506-0509; Departments of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky 40506-0509. Electronic address: kloconnor@uky.edu. LA - eng GR - R01 CA109136/CA/NCI NIH HHS/United States GR - T32 CA160003/CA/NCI NIH HHS/United States GR - T32 CA165990/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20150917 PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (AREG protein, human) RN - 0 (Amphiregulin) RN - 0 (EGF Family of Proteins) RN - 0 (EREG protein, human) RN - 0 (Epiregulin) RN - 0 (HGF protein, human) RN - 0 (Integrin alpha6beta4) RN - 0 (RNA, Messenger) RN - 0 (RNA, Neoplasm) RN - 67256-21-7 (Hepatocyte Growth Factor) RN - EC 2.7.10.1 (EGFR protein, human) RN - EC 2.7.10.1 (ErbB Receptors) RN - EC 3.4.24.7 (MMP1 protein, human) RN - EC 3.4.24.7 (Matrix Metalloproteinase 1) SB - IM MH - Amphiregulin MH - Cell Line, Tumor MH - Cell Movement MH - EGF Family of Proteins/genetics/metabolism MH - Epiregulin/genetics/metabolism MH - ErbB Receptors/antagonists & inhibitors/genetics/*metabolism MH - Gene Knockdown Techniques MH - Hepatocyte Growth Factor/*metabolism MH - Humans MH - Integrin alpha6beta4/genetics/*metabolism MH - Matrix Metalloproteinase 1/genetics/metabolism MH - Models, Biological MH - Neoplasm Invasiveness MH - Pancreatic Neoplasms/genetics/metabolism/pathology MH - RNA, Messenger/genetics/metabolism MH - RNA, Neoplasm/genetics/metabolism MH - Signal Transduction MH - Tumor Microenvironment MH - Up-Regulation PMC - PMC4646402 OTO - NOTNLM OT - pancreatic cancer OT - protein secretion OT - receptor protein-tyrosine kinase OT - receptor protein-tyrosine kinases OT - receptor regulation OT - receptor tyrosine kinase OT - signal transduction EDAT- 2015/09/19 06:00 MHDA- 2016/04/23 06:00 PMCR- 2016/11/06 CRDT- 2015/09/19 06:00 PHST- 2015/08/20 00:00 [received] PHST- 2015/09/19 06:00 [entrez] PHST- 2015/09/19 06:00 [pubmed] PHST- 2016/04/23 06:00 [medline] PHST- 2016/11/06 00:00 [pmc-release] AID - S0021-9258(20)49488-8 [pii] AID - M115.686873 [pii] AID - 10.1074/jbc.M115.686873 [doi] PST - ppublish SO - J Biol Chem. 2015 Nov 6;290(45):27228-27238. doi: 10.1074/jbc.M115.686873. Epub 2015 Sep 17.