Oncogenic K-ras and loss of smad4 mediate invasion by activating an EGFR/NF-?B Axis that induces expression of MMP9 and uPA in human pancreas progenitor cells

Alakesh Bera, Shujie Zhao, Lin Cao, Paul J. Chiao, James W. Freeman

Research output: Contribution to journalArticle

Abstract

Abstract Activating K-Ras mutations and inactivating mutations of Smad4 are two common genetic alterations that occur in the development and progression of pancreatic ductal adenocarcinomas (PDAC). To further study the individual and combinatorial roles of these two mutations in the pathogenesis of PDAC, immortalized human pancreas nestin postive cells (HPNE) were genetically modified by either expressing oncogenic K-Ras (HPNE/K-Ras), by shRNA knock down of Smad4 (HPNE/ShSmad4) or by creating both alterations in the same cell line (HPNE/K-Ras/ ShSmad4). We previously found that expression of oncogenic K-Ras caused an increase in expression of EGFR and loss of Smad4 further enhanced the up regulation in expression of EGFR and that this increase in EGFR was sufficient to induce invasion. Here we further investigated the mechanism that links mutational alterations and EGFR expression with invasion. The increase in EGFR signaling was associated with up regulation of MMP9 and uPA protein and activity. Moreover, the increase in EGFR signaling promoted a nuclear translocation and binding of RelA (p65), a subunit of NF-?B, to the promoters of both MMP-9 and uPA. Treatment of HPNE/K-Ras/ShSmad4 cells with an inhibitor of EGFR reduced EGF-mediated NF-?B nuclear translocation and inhibitors of either EGFR or NF-?B reduced the increase in MMP-9 or uPA expression. In conclusion, this study provides the mechanism of how a combination of oncogenic K-Ras and loss of Smad4 causes invasion and provides the basis for new strategies to inhibit metastases.

Original languageEnglish (US)
Article numbere82282
JournalPLoS One
Volume8
Issue number12
DOIs
StatePublished - Dec 5 2013
Externally publishedYes

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Nestin
pancreas
stem cells
Pancreas
Stem Cells
gelatinase B
Matrix Metalloproteinases
cells
adenocarcinoma
mutation
Mutation
Adenocarcinoma
Up-Regulation
human cell lines
Epidermal Growth Factor
Small Interfering RNA
metastasis
Cells
pathogenesis
promoter regions

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Oncogenic K-ras and loss of smad4 mediate invasion by activating an EGFR/NF-?B Axis that induces expression of MMP9 and uPA in human pancreas progenitor cells. / Bera, Alakesh; Zhao, Shujie; Cao, Lin; Chiao, Paul J.; Freeman, James W.

In: PLoS One, Vol. 8, No. 12, e82282, 05.12.2013.

Research output: Contribution to journalArticle

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abstract = "Abstract Activating K-Ras mutations and inactivating mutations of Smad4 are two common genetic alterations that occur in the development and progression of pancreatic ductal adenocarcinomas (PDAC). To further study the individual and combinatorial roles of these two mutations in the pathogenesis of PDAC, immortalized human pancreas nestin postive cells (HPNE) were genetically modified by either expressing oncogenic K-Ras (HPNE/K-Ras), by shRNA knock down of Smad4 (HPNE/ShSmad4) or by creating both alterations in the same cell line (HPNE/K-Ras/ ShSmad4). We previously found that expression of oncogenic K-Ras caused an increase in expression of EGFR and loss of Smad4 further enhanced the up regulation in expression of EGFR and that this increase in EGFR was sufficient to induce invasion. Here we further investigated the mechanism that links mutational alterations and EGFR expression with invasion. The increase in EGFR signaling was associated with up regulation of MMP9 and uPA protein and activity. Moreover, the increase in EGFR signaling promoted a nuclear translocation and binding of RelA (p65), a subunit of NF-?B, to the promoters of both MMP-9 and uPA. Treatment of HPNE/K-Ras/ShSmad4 cells with an inhibitor of EGFR reduced EGF-mediated NF-?B nuclear translocation and inhibitors of either EGFR or NF-?B reduced the increase in MMP-9 or uPA expression. In conclusion, this study provides the mechanism of how a combination of oncogenic K-Ras and loss of Smad4 causes invasion and provides the basis for new strategies to inhibit metastases.",
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