The anti‑epithelial cell adhesion molecule (EpCAM) monoclonal antibody EpMab‑16 exerts antitumor activity in a mouse model of colorectal adenocarcinoma

Hosono,Hideki; Ohishi,Tomokazu; Takei,Junko; Asano,Teizo; Sayama,Yusuke; Kawada,Manabu; Kaneko,Mika K.; Kato,Yukinari

doi:10.3892/ol.2020.12246

The anti‑epithelial cell adhesion molecule (EpCAM) monoclonal antibody EpMab‑16 exerts antitumor activity in a mouse model of colorectal adenocarcinoma

Authors:
- Hideki Hosono
- Tomokazu Ohishi
- Junko Takei
- Teizo Asano
- Yusuke Sayama
- Manabu Kawada
- Mika K. Kaneko
- Yukinari Kato
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Affiliations: Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Miyagi 980‑8575, Japan, Institute of Microbial Chemistry (BIKAKEN), Numazu, Microbial Chemistry Research Foundation, Numazu, Shizuoka 410‑0301, Japan
Published online on: October 23, 2020 https://doi.org/10.3892/ol.2020.12246
Article Number: 383
Copyright : © Hosono et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].

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Abstract

The epithelial cell adhesion molecule (EpCAM), which is a calcium‑independent homophilic intercellular adhesion factor, contributes to cell signaling, differentiation, proliferation and migration. EpCAM is essential for carcinogenesis in numerous types of human cancer. The purpose of the present study was to establish an anti‑EpCAM monoclonal antibody (mAb) for targeting colorectal adenocarcinomas. Thus, an anti‑EpCAM mAb, EpMab‑16 (IgG2a, κ), was established by immunizing mice with EpCAM‑overexpressing CHO‑K1 cells, and validated using flow cytometry, western blot, and immunohistochemical analyses. EpMab‑16 reacted with endogenous EpCAM specifically in a colorectal adenocarcinoma cell line as determined by flow cytometry and western blot analyses. Immunohistochemical analysis demonstrated that EpMab‑16 stained a plasma membrane‑like pattern in clinical colorectal adenocarcinoma tissues. The dissociation constant (K_D) for EpMab‑16 in a Caco‑2 colorectal adenocarcinoma cell line determined by flow cytometry was 1.8x10^‑8 M, suggesting moderate binding affinity of EpMab‑16 for EpCAM. Whether the EpMab‑16 induced antibody‑dependent cellular cytotoxicity (ADCC) and complement‑dependent cytotoxicity (CDC) against Caco‑2 or antitumor activity was then assessed in a murine xenograft model. In vitro experiments revealed strong ADCC and CDC induction in Caco‑2 cells by EpMab‑16 treatment. In vivo experiments in a Caco‑2 xenograft model demonstrated that EpMab‑16 treatment significantly reduced tumor growth compared with that in mice treated with the control mouse IgG. These results suggested that EpMab‑16 may be a promising treatment option for EpCAM‑expressing colorectal adenocarcinomas.

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