Abstract

A subset of patients with Ewing's sarcoma responds to anti-insulin-like growth factor-1 receptor (IGF-1R) antibodies. Mechanisms of sensitivity and resistance are unknown. We investigated whether an anti-IGF-1R antibody acts via a pathway that could also be suppressed by small interfering (si) RNA against the EWS/FLI-1 fusion protein, the hallmark of Ewing's sarcoma. The growth of two Ewing's sarcoma cell lines (TC-32 and TC-71) was inhibited by the fully human anti-IGF-1R antibody, R1507 (clonogenic and MTT assays). TC-32 and TC-71 cells express high levels of IGF-2, while RD-ES and A4573 Ewing's cell lines, which were less responsive to R1507 in our assays, express low or undetectable IGF-2, respectively. TC-71 cells also expressed high levels of IGF-1R, and R1507 decreased steady-state levels of this receptor by internalization/degradation, an effect which was associated with a decrease in p-IGF-1R, p-IRS-1, and p-Akt. EWS/FLI-1 siRNA also decreased p-Akt, due to its ability to increase IGF-BP3 levels and subsequently decrease IGF-1 and IGF-2 levels, thus inhibiting signaling through p-IGF-1R. This inhibition correlated with growth suppression and apoptosis. The attenuation of Akt activation was confirmed in TC-71 and HEK-293 (human embryonic kidney) cells by transfecting them with IGF-1R siRNA. We conclude that antibodies and siRNA to IGF-1R, as well as siRNA to EWS/FLI-1, act via intersecting IGF/IGF-1R signals that suppress a common point in this pathway, namely the phosphorylation of Akt.

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Authors

Huang HJ, Angelo LS, Rodon J, Sun M, Kuenkele KP, Parsons HA, Trent JC, Kurzrock R

Institution

Phase I Program, Department of Investigational Cancer Therapeutics, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, United States of America.

Source

PloS one 6:10 2011 pg e26060

MeSH

Antibodies, Monoclonal
Apoptosis
Cell Line, Tumor
Cell Proliferation
Colony-Forming Units Assay
Down-Regulation
HEK293 Cells
Humans
Insulin Receptor Substrate Proteins
Insulin-Like Growth Factor Binding Protein 3
Insulin-Like Growth Factor II
Oncogene Proteins, Fusion
Phosphorylation
Polymorphism, Genetic
Protein Structure, Tertiary
Proto-Oncogene Protein c-fli-1
Proto-Oncogene Proteins c-akt
RNA, Small Interfering
RNA-Binding Protein EWS
Receptor, IGF Type 1
Receptor, IGF Type 2
Reproducibility of Results
Sarcoma, Ewing
Signal Transduction
Somatomedins
Transfection

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

22022506