TY - JOUR
T1 - Downregulation of Nodal inhibits metastatic progression in retinoblastoma
AU - Asnaghi, Laura
AU - White, David T.
AU - Yoon, Lynn
AU - Price, Antoinette
AU - Lee, Grace Y.
AU - Sahoo, Arpan
AU - Mumm, Jeff S.
AU - Eberhart, Charles G.
N1 - Funding Information:
This research was supported by the NIH Grant R21CA229919 (to L.A. and C.G.E.), the core grant EY001765, and The Jenny Fund.
Funding Information:
This study was supported by the NIH Grant R21CA229919 (to L.A. and C.G.E.), the core grant EY001765, and The Jenny Fund. Dr. Laura Asnaghi is currently employed at the National Institutes of Health. This article was prepared while Dr. Asnaghi was employed at the Johns Hopkins University. The opinions expressed in this article are the author's own and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government.
PY - 2019/8/26
Y1 - 2019/8/26
N2 - Retinoblastoma is the most common intraocular malignancy in children. We previously found that the ACVR1C/SMAD2 pathway is significantly upregulated in invasive retinoblastoma samples from patients. Here we studied the role of an ACVR1C ligand, Nodal, in regulating growth and metastatic dissemination in retinoblastoma. Inhibition of Nodal using multiple short hairpin (shRNAs) in WERI Rb1 and Y79 retinoblastoma cell cultures reduced growth by more than 90%, as determined by CCK-8 growth assay. Proliferation was also significantly inhibited, as found by Ki67 assay. These effects were paralleled by inhibition in the phosphorylation of the downstream effector SMAD2, as well as induction of apoptosis, as we observed more than three-fold increase in the percentage of cells positive for cleaved-caspase-3 or expressing cleaved-PARP1. Importantly, we found that downregulation of Nodal potently suppressed invasion in vitro, by 50 to 80%, as determined by transwell invasion assay (p = 0.02). Using an orthotopic model of retinoblastoma in zebrafish, we found 34% reduction in the ability of the cells to disseminate outside the eye, when Nodal was knocked down by shRNA (p = 0.0003). These data suggest that Nodal plays an important role in promoting growth, proliferation and invasion in retinoblastoma, and can be considered a new therapeutic target for both primary tumor growth and metastatic progression.
AB - Retinoblastoma is the most common intraocular malignancy in children. We previously found that the ACVR1C/SMAD2 pathway is significantly upregulated in invasive retinoblastoma samples from patients. Here we studied the role of an ACVR1C ligand, Nodal, in regulating growth and metastatic dissemination in retinoblastoma. Inhibition of Nodal using multiple short hairpin (shRNAs) in WERI Rb1 and Y79 retinoblastoma cell cultures reduced growth by more than 90%, as determined by CCK-8 growth assay. Proliferation was also significantly inhibited, as found by Ki67 assay. These effects were paralleled by inhibition in the phosphorylation of the downstream effector SMAD2, as well as induction of apoptosis, as we observed more than three-fold increase in the percentage of cells positive for cleaved-caspase-3 or expressing cleaved-PARP1. Importantly, we found that downregulation of Nodal potently suppressed invasion in vitro, by 50 to 80%, as determined by transwell invasion assay (p = 0.02). Using an orthotopic model of retinoblastoma in zebrafish, we found 34% reduction in the ability of the cells to disseminate outside the eye, when Nodal was knocked down by shRNA (p = 0.0003). These data suggest that Nodal plays an important role in promoting growth, proliferation and invasion in retinoblastoma, and can be considered a new therapeutic target for both primary tumor growth and metastatic progression.
KW - Invasion
KW - Nodal
KW - Proliferation
KW - Retinoblastoma
UR - http://www.scopus.com/inward/record.url?scp=85071646812&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85071646812&partnerID=8YFLogxK
U2 - 10.1186/s40478-019-0785-4
DO - 10.1186/s40478-019-0785-4
M3 - Article
C2 - 31451106
AN - SCOPUS:85071646812
SN - 2051-5960
VL - 7
SP - 137
JO - Acta Neuropathologica Communications
JF - Acta Neuropathologica Communications
IS - 1
ER -