J Korean Assoc Oral Maxillofac Surg.  2010 Jun;36(3):177-185. 10.5125/jkaoms.2010.36.3.177.

Anti-epidermal growth factor receptor (EGFR) monoclonal antibody and DNA topoisomerase inhibitor reduce growth of salivary adenoid cystic carcinoma in a murine model

• Park YW
• Lee HS

• KMID: 2189945
• DOI: http://doi.org/10.5125/jkaoms.2010.36.3.177

Abstract

INTRODUCTION
Epidermal growth factor receptor (EGFR) is expressed in human epithelial tumors including salivary cancers, and known to be correlated with tumor progression and poor clinical courses in some epithelial tumors. In this study, we determined the therapeutic effect of the anti-EGFR monoclonal antibody Erbitux (C225, cetuximab) in combination with the DNA topoisomerase I inhibitor irinotecan (CPT-11) on human salivary adenoid cystic carcinoma (SACC) cells growing in nude mice.
MATERIALS AND METHODS
At first, immunocytochemical analysis for the expression of EGFR and phosphorylated EGFR (pEGFR) on a human salivary ACC cell line (ACC3). To determine the in vivo effects of Erbitux and CPT-11, nude mice with orthotopic parotid tumors were randomized to receive intraperitoneal Erbitux (1 mg) two times per week, intraperitoneal Irinotecan (50 mg/kg) once per week, Erbitux plus CPT-11, or placebo. (control) Tumor volume and weight were measured. And mechanisms of in vivo activity of Erbitux and/or CPT-11 were determined by immunohistochemical/immunofluorescent analyses.
RESULTS
Immunocytochemical staining of ACC3 demonstrated that EGFR was expressed and phosphorylated. CPT-11 inhibited ACC tumor growth in nude mice. Tumors of mice treated with CPT-11 and CPT-11 plus Erbitux exhibited increased tumor cell apoptosis and decreased microvessel density, which correlated with a decrease in the tumor volume in nude mice. But, CPT-11 seems not to be synergistic with Erbitux in our ACC3 model system.
CONCLUSION
These results suggest that anti-EGFR monoclonal antibody and the DNA topoisomerase I inhibitor will be effective in the treatment of recurred or metastatic lesions of salivary ACC.

Keyword

Monoclonal antibodies; Epidermal growth factor receptor; DNA topoisomerases type 1; Adenoid cystic carcinoma; Salivary gland neoplasms; Parotid neoplasms

MeSH Terms

Adenoids
Animals
Antibodies, Monoclonal
Antibodies, Monoclonal, Humanized
Apoptosis
Camptothecin
Carcinoma, Adenoid Cystic
Cell Line
Cetuximab
DNA
DNA Topoisomerases, Type I
Humans
Mice
Mice, Nude
Microvessels
Parotid Neoplasms
Receptor, Epidermal Growth Factor
Salivary Gland Neoplasms
Tumor Burden
Antibodies, Monoclonal
Antibodies, Monoclonal, Humanized
Camptothecin
DNA
DNA Topoisomerases, Type I
Receptor, Epidermal Growth Factor

Figure

• 

  Fig. 1. Immunocytochemical analysis of the chamber slides in salivary adenoid cystic carcinoma cells. ACC3 cells were immunostained for the expression of EGFR and phosphorylated EGFR. Representative results are shown.(original magnification ×200) (ACC3: adenoid cystic carcinoma cell line, EGFR: epidermal growth factor receptor)

• 

  Fig. 2. A. CPT-11 inhibits the growth of salivary ACC xenografts in nude mice. ACC cells (5×105) were injected into the parotid gland of nude mice. Seven days after the injection of the tumor cell suspension, the mice (n=10) were treated with intraperitoneal injection of Erbitux at 1 mg/injection twice weekly, CPT-11 given via intraperitoneal injection at 50 mg/kg once a week, or both Erbitux and CPT-11. Ponits, mean tumor volume measured twice per week and expressed once per week.(bars: SE) B. Tumor weights of each group at the time of sacrifice. Results are means±SE. (ACC: adenoid cystic carcinoma)

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  Fig. 3. Representative gross features of the tumors of each group at the time of sacrifice. A. Control B. Erbitux 1 mg/injection, twice weekly. C. CPT-11 50 mg/kg, once a week. D. CPT-11+Erbitux.

• 

  Fig. 4. Histological (H&E staining), immunohistochemical/immunofluorescent and TUNEL analyses for degree of EGFR tyrosine kinase phosphorylation, cell proliferation, and apoptosis. After 5 weeks of treatment with Erbitux, CPT-11, or Erbitux+CPT11, ACC3 orthotopic tumors were sectioned and immunostained for EGFR, pEGFR, PCNA, and stained with TUNEL. Treatment with Erbitux alone or in combination with CPT-11, inhibited the degree of phosphorylation of EGFR. Note that all the treatment group showed decreased number of PCNA-positive tumor cells. And Some of CPT11-treated tumor cells demonstrated apoptosis. Repersentative features are shown.(original magnification ×100, in panel of TUNEL ×200) (EGFR: epidermal growth factor receptor, pEGFR: phosphorylated EGFR, ACC3: adenoid cystic carcinoma cell line, PCNA: proliferating cell nuclear antigen, TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling)

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  Fig. 5. Immunohistochemical and immunofluorescent double staining for tumor-associated endothelial cells in Erbitux/CPT-11-treated orthotopic tumors of ACC3. Microvessel density was decreased in CPT-11-treated tumors, but there was no apoptosis of the tumor-associated endothelial cells.(original magnification ×100, in panel of CD31 ×200, in panel of CD31/TUNEL ×200) (ACC3: adenoid cystic carcinoma cell line)

Reference

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