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01-09-2012 | Original article

Cellular and molecular properties of ⁹⁰Y-labeled cetuximab in combination with radiotherapy on human tumor cells in vitro

Authors: M. Saki, M. Toulany, W. Sihver, M. Zenker, J.-M. Heldt, B. Mosch, H.-J. Pietzsch, M. Baumann, J. Steinbach, Prof. Dr. H.P. Rodemann

Published in: Strahlentherapie und Onkologie | Issue 9/2012

Abstract

Purpose

Anti-EGFR antibody cetuximab (C225) is used in combination with radiotherapy of head and neck squamous cell carcinoma (HNSCC) patients. We investigated whether conjugation of cetuximab with trans-cyclohexyl-diethylene-triamine-pentaacetic acid (CHX-A″-DTPA) and radiolabeling with ⁹⁰Yttrium affect the molecular and cellular function of cetuximab and improve its combined effect with external-beam irradiation (EBI).

Methods

The following cell lines were used: HNSCC UT5, SAS, FaDu, as well as A43, Chinese hamster ovary cells (CHO), and human skin fibroblast HSF7. Binding affinity and kinetics, specificity, retention, and the combination of ⁹⁰Y-cetuximab with EBI were evaluated.

Results

Control cetuximab and CHX-A″-DTPA-cetuximab blocked the proliferation activity of UT5 cells. In combination with EBI, CHX-A″-DTPA-cetuximab increased the radiosensitivity of UT5 to a similar degree as control cetuximab did. In contrast, in SAS and HSF7 cells neither proliferation nor radiosensitivity was affected by either of the antibodies. Binding [⁹⁰Y]Y-CHX-A″-DTPA-cetuximab (⁹⁰Y-cetuximab) to EGFR in HNSCC cells occurred time dependently with a maximum binding at 24 h. Retention of ⁹⁰Y-cetuximab was similar in both HNSCC cell lines; 24 h after treatment, approximately 90% of bound activity remained in the cell layer. Competition assays, using cell membranes in the absence of an internalized fraction of cetuximab, showed that the cetuximab affinity is not lost as a result of conjugation with CHX-A″-DTPA. Cetuximab and CHX-A″-DTPA-cetuximab blocked EGF-induced Y1068 phosphorylation of EGFR. The lack of an effect of cetuximab on EGF-induced Akt and ERK1/2 phosphorylation and the inhibition of irradiation (IR)-induced Akt and ERK1/2 phosphorylation by cetuximab were not affected by DTPA conjugation. ⁹⁰Y-cetuximab in combination with EBI resulted in a pronounced inhibition of colony formation of HNSCC cells.

Conclusions

Conjugation of CHX-A″-DTPA to cetuximab does not alter the cellular and biological function of cetuximab. ⁹⁰Y-labeling of cetuximab in combination with EBI may improve radiotherapy outcome.

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Title: Cellular and molecular properties of 90Y-labeled cetuximab in combination with radiotherapy on human tumor cells in vitro
Authors: M. Saki
M. Toulany
W. Sihver
M. Zenker
J.-M. Heldt
B. Mosch
H.-J. Pietzsch
M. Baumann
J. Steinbach
Prof. Dr. H.P. Rodemann
Publication date: 01-09-2012
Publisher: Springer-Verlag
Published in: Strahlentherapie und Onkologie / Issue 9/2012
Print ISSN: 0179-7158
Electronic ISSN: 1439-099X
DOI: https://doi.org/10.1007/s00066-012-0121-4

At a glance: The STEP trials

Springer Medicine

Cellular and molecular properties of ⁹⁰Y-labeled cetuximab in combination with radiotherapy on human tumor cells in vitro

Abstract

Purpose

Methods

Results

Conclusions

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

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