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First published online August 14, 2008, 10.2967/jnumed.108.052316
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Correlating EGFR Expression with Receptor-Binding Properties and Internalization of 64Cu-DOTA-Cetuximab in 5 Cervical Cancer Cell Lines

Martin Eiblmaier1, Laura A. Meyer1, Mark A. Watson2, Paula M. Fracasso3, Linda J. Pike4 and Carolyn J. Anderson1,5

1 Mallinckrodt Institute of Radiology, Washington University School of Medicine, St. Louis, Missouri; 2 Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri; 3 Department of Medicine, Washington University School of Medicine, St. Louis, Missouri; 4 Department of Biochemistry, Washington University School of Medicine, St. Louis, Missouri; and 5 Department of Chemistry, Washington University School of Medicine, St. Louis, Missouri


Figure 1
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  		FIGURE 1.  Transcriptional profiling of cervical cancer cell lines with U133Plus2 GeneChip microarrays (Affymetrix). EGFR expression is based on 4 different oligonucleotide probe sets (each column shows response to 1 probe set). Cell lines are ordered based on their relative levels of EGFR probe signals. Cell lines with suffix designation "-a" were grown and harvested independently.

 

Figure 2
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  		FIGURE 2.  Saturation receptor binding of 64Cu-DOTA-cetuximab to 5 cervical cancer cell lines (n = 3). EGFR densities (Bmax in fmol/mg) on cell surface were, from high to low, CaSki ({blacksquare}, 2,130 ± 60) > ME-180 ({Delta}, 820 ± 40) > DoTc2 4510 (•, 610 ± 20) > HeLa ({diamond}, 310 ± 110) > C-33A ({blacktriangledown}, not applicable).

 

Figure 3
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  		FIGURE 3.  Internalization of 64Cu-DOTA-cetuximab (n = 3). (A) Internalization standardized to protein mass over 4 h displayed same relative order as seen by expression profiling and receptor binding: CaSki ({blacksquare}) > ME-180 ({Delta}) > DoTc2 4510 (•) > HeLa ({diamond}). (B) Rate of internalization in first 15 min of exposure to 64Cu-DOTA-cetuximab, displayed as internalized to surface-bound activity.

 

Figure 4
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  		FIGURE 4.  Biodistribution of 64Cu-DOTA-cetuximab in CaSki tumors implanted into SCID mice at 24 h after injection.

 

Figure 5
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  		FIGURE 5.  Small-animal PET imaging of 64Cu-DOTA-cetuximab in CaSki tumor–bearing SCID mice at 24 h after injection: coronal (A), sagittal (B), and transaxial (C) views.

 




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