title: Pertuzumab in combination with trastuzumab shows significantly enhanced antitumor activity in HER2-positive human gastric cancer xenograft models.
abstract: OBJECTIVE
  We investigated the antitumor activity of the combination of two different humanized monoclonal human epidermal growth factor receptor (HER) 2 antibodies, pertuzumab and trastuzumab, for gastric cancer.
  
  
  METHODS
  Tumor mouse xenograft models were used to examine antitumor activity. Cell proliferation was examined using crystal violet staining. HER family proteins' expression was analyzed by ELISA and immunohistochemistry. Phosphorylated proteins and heterodimers were detected by Western blotting and in situ proximity ligation assay (PLA), respectively. Apoptosis activity was examined by caspase 3/7 activity. Antibody-dependent cellular cytotoxicity (ADCC) activity was detected by xCELLigence. Microvessel density was examined by CD31 staining.
  
  
  RESULTS
  Pertuzumab in combination with trastuzumab showed significant antitumor activity compared with each monotherapy in NCI-N87, an HER2-positive human gastric cancer xenograft model. The efficacy was stronger than that of the maximum effective dose with each monotherapy. Similar antitumor activity was shown in 4-1ST, another HER2-positive gastric cancer model, but not in MKN-28, an HER2-negative model. Combining pertuzumab with trastuzumab enhanced cell growth inhibition and apoptosis activity by inhibiting EGFR-HER2 heterodimerization and the phosphorylation of these receptors and their downstream factors. This effect was also seen in HER2-HER3 signaling. Furthermore, pertuzumab in combination with trastuzumab potentiated the ADCC activity of those antibodies and reduced tumor microvessel density.
  
  
  CONCLUSIONS
  We showed the significantly enhanced efficacy of pertuzumab combining with trastuzumab for HER2 overexpressing gastric cancer through the potentiation of cell growth inhibition, apoptosis activity, cell killing activity by ADCC, and antiangiogenic activity. This study suggests the clinical benefit of combination therapy with pertuzumab and trastuzumab for patients with HER2-positive gastric cancers.
authors.affiliation: Product Research Department, API Process Development Department, and Discovery Platform Technology Department, Chugai Pharmaceutical Co., Kanagawa, Japan.
authors.forename: Yoriko
authors.initials: Y
authors.lastname: Yamashita-Kashima
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Iijima
authors.forename: Keigo
authors.initials: K
authors.lastname: Yorozu
authors.forename: Koh
authors.initials: K
authors.lastname: Furugaki
authors.forename: Mitsue
authors.initials: M
authors.lastname: Kurasawa
authors.forename: Masateru
authors.initials: M
authors.lastname: Ohta
authors.forename: Kaori
authors.initials: K
authors.lastname: Fujimoto-Ouchi
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D066246:ErbB Receptors / Q000276:immunology*; D018734:Genes, erbB-2; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D015398:Signal Transduction / Q000187:drug effects; D013274:Stomach Neoplasms / Q000188:drug therapy* / Q000235:genetics*; D000068878:Trastuzumab; D023041:Xenograft Model Antitumor Assays
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D066246:ErbB Receptors; C485206:pertuzumab; D000068878:Trastuzumab