TY - JOUR
T1 - Extracellular superoxide dismutase inhibits hepatocyte growth factor-mediated breast cancer-fibroblast interactions
AU - Golden, Briana Ormsbee
AU - Griess, Brandon
AU - Mir, Shakeel
AU - Fitzgerald, Matthew
AU - Kuperwasser, Charlotte
AU - Domann, Frederick
AU - Teoh-Fitzgerald, Melissa
N1 - Funding Information:
This work was financially supported by grants
Funding Information:
We would like to thank Michelle Desler for her technical support with some of the cell culture work. We are grateful for the kind gift of the anti-EcSOD antibody and MnTE-2-PyP from Dr. James Crapo (National Jewish Medical and Research Center, Denver, CO, USA). This work was financially supported by grants from the NIH R01 CA115438 (FE Domann), NIH R0ICA182086A (Teoh-Fitzgerald), Redox Biology Pilot Project Fund (NTSBRDF, Uni. of Nebraska, Lincoln) (Teoh-Fitzgerald), and Nebraska Department of Health and Human Services Award LB506 (Teoh-Fitzgerald). Brandon Griess was supported by the Eppley Institute in Cancer Biology Training Grant (NCI T32CA009476). EPR spectroscopy data were collected in the University of Nebraska EPR Spectroscopy Core, which is supported in part, by the National Institute of General Medical Sciences of the National Institute of Health (P30GM103335) awarded to the University of Nebraska's Redox Biology Center.
Publisher Copyright:
© Ormsbee Golden et al.
PY - 2017
Y1 - 2017
N2 - We have previously shown tumor suppressive effects of extracellular superoxide dismutase, EcSOD in breast cancer cells. In this study, an RTK signaling array revealed an inhibitory effect of EcSOD on c-Met phosphorylation and its downstream kinase c-Abl in MDA-MB231 cells. Moreover, an extracellular protein array showed that thrombospondin 1 (TSP-1), a scavenger of the c-Met ligand, hepatocyte growth factor (HGF) is significantly up-regulated in EcSOD overexpressing cells (Ec.20). We further determined the effects of EcSOD on HGF/c-Met-mediated cancer-fibroblast interactions by co-culturing normal fibroblasts (RMF) or RMF which overexpresses HGF (RMF-HGF) with MDA-MB231 cells. We observed that while RMF-HGF significantly promoted Matrigel growth of MDA-MB231, overexpression of EcSOD inhibited the HGF-stimulated growth. Similarly, a SOD mimetic, MnTE-2-PyP, inhibited HGF-induced growth and invasion of MDA-MB231. In addition, a long-term heterotypic co-culture study not only showed that Ec.20 cells are resistant to RMF-HGF-induced invasive stimulation but RMF-HGF that were co-cultured with Ec.20 cells showed an attenuated phenotype, suggesting an oxidative-mediated reciprocal interaction between the two cell types. In addition, we demonstrated that RMF-HGF showed an up-regulation of an ROS-generating enzyme, NADPH oxidase 4 (Nox4). Targeting this pro-oxidant significantly suppressed the activated phenotype of RMF-HGF in a collagen contraction assay, suggesting that RMF-HGF contributes to the oxidative tumor microenvironment. We have further shown that scavenging ROS with EcSOD significantly inhibited RMFHGF- stimulated orthotopic tumor growth of MDA-MB231. This study suggests the loss of EcSOD in breast cancer plays a pivotal role in promoting the HGF/c-Met-mediated cancer-fibroblast interactions.
AB - We have previously shown tumor suppressive effects of extracellular superoxide dismutase, EcSOD in breast cancer cells. In this study, an RTK signaling array revealed an inhibitory effect of EcSOD on c-Met phosphorylation and its downstream kinase c-Abl in MDA-MB231 cells. Moreover, an extracellular protein array showed that thrombospondin 1 (TSP-1), a scavenger of the c-Met ligand, hepatocyte growth factor (HGF) is significantly up-regulated in EcSOD overexpressing cells (Ec.20). We further determined the effects of EcSOD on HGF/c-Met-mediated cancer-fibroblast interactions by co-culturing normal fibroblasts (RMF) or RMF which overexpresses HGF (RMF-HGF) with MDA-MB231 cells. We observed that while RMF-HGF significantly promoted Matrigel growth of MDA-MB231, overexpression of EcSOD inhibited the HGF-stimulated growth. Similarly, a SOD mimetic, MnTE-2-PyP, inhibited HGF-induced growth and invasion of MDA-MB231. In addition, a long-term heterotypic co-culture study not only showed that Ec.20 cells are resistant to RMF-HGF-induced invasive stimulation but RMF-HGF that were co-cultured with Ec.20 cells showed an attenuated phenotype, suggesting an oxidative-mediated reciprocal interaction between the two cell types. In addition, we demonstrated that RMF-HGF showed an up-regulation of an ROS-generating enzyme, NADPH oxidase 4 (Nox4). Targeting this pro-oxidant significantly suppressed the activated phenotype of RMF-HGF in a collagen contraction assay, suggesting that RMF-HGF contributes to the oxidative tumor microenvironment. We have further shown that scavenging ROS with EcSOD significantly inhibited RMFHGF- stimulated orthotopic tumor growth of MDA-MB231. This study suggests the loss of EcSOD in breast cancer plays a pivotal role in promoting the HGF/c-Met-mediated cancer-fibroblast interactions.
KW - Breast cancer
KW - C-Met
KW - Cancer-associated fibroblasts
KW - EcSOD
KW - Tumor-stroma interactions
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U2 - 10.18632/oncotarget.22379
DO - 10.18632/oncotarget.22379
M3 - Article
C2 - 29296173
AN - SCOPUS:85037367942
VL - 8
SP - 107390
EP - 107408
JO - Oncotarget
JF - Oncotarget
SN - 1949-2553
IS - 64
ER -