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Constitutive K‐RasG12D Activation of ERK2 Specifically Regulates 3D Invasion of Human Pancreatic Cancer Cells via MMP‐1
Author(s) -
Botta Gregory P,
Reginato Mauricio J,
Rustgi Anil K,
Lelkes Peter I
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.975.1
Subject(s) - invadopodia , cancer research , stromal cell , pancreatic cancer , gene silencing , small hairpin rna , biology , matrix metalloproteinase , cancer , metastasis , cell culture , gene knockdown , gene , genetics
Pancreatic ductal adenocarcinomas (PDAC) are highly invasive & metastatic neoplasms unresponsive to current therapies. Overwhelmingly, PDAC harbors constitutive, oncogenic mutations in K‐ Ras G12D that exist prior to invasion. Histologic & genetic analyses of human PDAC exhibit increased expression of ERK1/2 and pro‐invasive MMPs; indicators of poor prognosis. However, the distinct molecular mechanisms necessary for K‐ Ras – ERK1/2 signaling & its influence on MMP‐directed stromal invasion in primary human pancreatic ductal epithelial cells (PDECs) has yet to be elucidated in 3D. Expression of oncogenic K‐ Ras G12D alone in genetically‐defined PDECs reveals increased invadopodia & epithelial‐to‐mesenchymal transition markers, but only when cultured in a 3D basement membrane (BM) model. Activation of ERK2, but not ERK1, occurs only in K‐ Ras G12D mutated PDECs cultured in 3D & is necessary for invasion based upon pharmacologic & shRNA inhibition. Increased invasion of K‐ Ras G12D PDECs through the BM is associated with a specific microarray gene signature & induction of MMPs. Specifically, MMP‐1 RNA, protein, & proteolysis are amplified in K‐ Ras G12D PDECs when assayed by RT‐PCR, ELISA, & FRET. Importantly, MMP‐1 silencing mimics ERK2 inhibition & disrupts vertical invasion. ERK2‐isoform & MMP‐1 targeting in 3D are shown to be viable strategies to attenuate invasion of K‐ Ras G12D mutated human pancreatic cancer cells. GPB is funded by an F30 DK088402‐01.