Cilengitide and Cetuximab Reduce Cytokine Production and Colony Formation of Head and Neck Squamous Cell Carcinoma CellsEx Vivo.
To analyze ex vivo effects of combined targeting of the epidermal growth factor-receptor (EGFR) by cetuximab (E) plus αVβ3 and αVβ5 integrins by cilengitide (Cil) on colony formation of epithelial cells (CFec) and release of pro-angiogenetic and pro-inflammatory cytokines in head and neck squamous cell carcinoma (HNSCC) cells.



Collagenase-IV digests of 43 histopathological confirmed HNSCC cases were seeded into laminin-coated 96-well plates containing E, Cil, or Cil+E in final concentrations of 66.7 μg/ml, 10 μM, and 10 μM+66.7 μg/ml, respectively. Following the FLAVINO-assay protocol, supernatants were harvested after 3 days and adherent cells fixed in ethanol. Counting of CFecwas facilitated by FITC-labeled pan-cytokeratin antibodies. Out of 43 HNSCC cases, 39 had adherent growth (mean CFec≥4/well in triplicate controls). Cytokines in supernatants were measured using ELISA were interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1) and vascular endothelial growth factor A (VEGFA).



CFecon laminin was significantly reduced by Cil, E, and Cil+E. Cytokine measurements also revealed significant suppression of MCP-1, IL-6 and VEGFA. The strongest suppression of CFec, MCP-1 and VEGFA release was exerted by Cil and E combined. Efficacy of Cil+E exceeded those of the solely applied pharmaceutics but failed regarding significant synergism of both treatments as E was unable to significantly boost the effects of Cil. In contrast, IL-6 release was significantly suppressed by E but not by Cil, while their combination strongly reduced it.



Combined targeting of EGFR and integrins with E and Cil heightens their suppressive effects regarding CFecas well as release of pro-angiogenetic and pro-inflammatory cytokines.
Department of Otolaryngology, Head and Neck Surgery, University of Leipzig, Leipzig, Germany gunnar.wichmann@medizin.uni-leipzig.de.
Gunnar
G
Wichmann
Department of Otolaryngology, Head and Neck Surgery, University of Leipzig, Leipzig, Germany.
Susan
S
Cedra
Department of Cranio-Maxillo-Facial Surgery, Paracelsus Medical University Nuremberg Hospital South, Nuremberg, Germany.
Daphne
D
Schlegel
Department of Otolaryngology, Head and Neck Surgery, University of Leipzig, Leipzig, Germany.
Marlen
M
Kolb
Department of Otolaryngology, Head and Neck Surgery, University of Leipzig, Leipzig, Germany.
Susanne
S
Wiegand
Department of Otolaryngology, Head and Neck Surgery, University of Leipzig, Leipzig, Germany.
Andreas
A
Boehm
Department of Otolaryngology, Head and Neck Surgery, University of Leipzig, Leipzig, Germany.
Mathias
M
Hofer
Department of Otolaryngology, Head and Neck Surgery, University of Leipzig, Leipzig, Germany.
Andreas
A
Dietz
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy* / Q000276:immunology* / Q000473:pathology; D000068818:Cetuximab / Q000008:administration & dosage / Q000494:pharmacology*; D018932:Chemokine CCL2 / Q000096:biosynthesis; D016207:Cytokines / Q000096:biosynthesis*; D004847:Epithelial Cells / Q000187:drug effects / Q000473:pathology; D005260:Female; D006258:Head and Neck Neoplasms / Q000188:drug therapy* / Q000276:immunology* / Q000473:pathology; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis; D008297:Male; D008875:Middle Aged; D012910:Snake Venoms / Q000008:administration & dosage / Q000494:pharmacology*; D000077195:Squamous Cell Carcinoma of Head and Neck; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis
C111835:CCL2 protein, human; D018932:Chemokine CCL2; D016207:Cytokines; D015850:Interleukin-6; D012910:Snake Venoms; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C422910:Cilengitide; D000068818:Cetuximab
Head and neck cancer; cetuximab; chemoresponse ex vivo; cilengitide; epidermal growth factor-receptor; head and neck squamous cell carcinoma; integrin; predictive assay; targeted therapy