Overexpression of neuropilin-1 promotes constitutive MAPK signalling and chemoresistance in pancreatic cancer cells.
Neuropilin-1 (NRP-1) is a novel co-receptor for vascular endothelial growth factor (VEGF). Neuropilin-1 is expressed in pancreatic cancer, but not in nonmalignant pancreatic tissue. We hypothesised that NRP-1 expression by pancreatic cancer cells contributes to the malignant phenotype. To determine the role of NRP-1 in pancreatic cancer, NRP-1 was stably transfected into the human pancreatic cancer cell line FG. Signal transduction was assessed by Western blot analysis. Susceptibility to anoikis (detachment induced apoptosis) was evaluated by colony formation after growth in suspension. Chemosensitivity to gemcitabine or 5-fluorouracil (5-FU) was assessed by MTT assay in pancreatic cancer cells following NRP-1 overexpression or siRNA-induced downregulation of NRP-1. Differential expression of apoptosis-related genes was determined by gene array and further evaluated by Western blot analysis. Neuropilin-1 overexpression increased constitutive mitogen activated protein kinase (MAPK) signalling, possibly via an autocrine loop. Neuropilin-1 overexpression in FG cells enhanced anoikis resistance and increased survival of cells by > 30% after exposure to clinically relevant levels of gemcitabine and 5-FU. In contrast, downregulation of NRP-1 expression in Panc-1 cells markedly increased chemosensitivity, inducing > 50% more cell death at clinically relevant concentrations of gemcitabine. Neuropilin-1 overexpression also increased expression of the antiapoptotic regulator, MCL-1. Neuropilin-1 overexpression in pancreatic cancer cell lines is associated with (a) increased constitutive MAPK signalling, (b) inhibition of anoikis, and (c) chemoresistance. Targeting NRP-1 in pancreatic cancer cells may downregulate survival signalling pathways and increase sensitivity to chemotherapy.
Wey
J S
JS
Department of Surgical Oncology, Unit 444, The University of Texas, MD Anderson Cancer Center, PO Box 301402, Houston, TX 77230-1402, USA.
Gray
M J
MJ
Fan
F
F
Belcheva
A
A
McCarty
M F
MF
Stoeltzing
O
O
Somcio
R
R
Liu
W
W
Evans
D B
DB
Klagsbrun
M
M
Gallick
G E
GE
Ellis
L M
LM
D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D017209:Apoptosis; D015153:Blotting, Western; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm; D005472:Fluorouracil / Q000494:pharmacology*; D020869:Gene Expression Profiling; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D039942:Neuropilin-1 / Q000096:biosynthesis* / Q000502:physiology*; D010190:Pancreatic Neoplasms / Q000473:pathology*; D010641:Phenotype; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured; D015854:Up-Regulation; D000093542:Gemcitabine
D018906:Antineoplastic Agents, Alkylating; D003841:Deoxycytidine; D039942:Neuropilin-1; D020928:Mitogen-Activated Protein Kinases; D005472:Fluorouracil; D000093542:Gemcitabine