A novel human Fab antibody for Trop2 inhibits breast cancer growth in vitro and in vivo
Hong Lin
Huadong Medical Institute of Biotechniques, Nanjing, China
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Department of Otolaryngology-Head and Neck Surgery, Jiangsu Provincial Hospital, Nanjing, China
Jiangsu Provincial Blood Center, Nanjing, China
Search for more papers by this authorHuiling Zhang
Department of Gynecology and Obstetrics, Nanjing Maternal and Children Care Hospital Affiliated to Nanjing Medical University, Nanjing, China
Search for more papers by this authorJun Wang
Department of Hematology and Oncology, Jiangsu Provincial Hospital, Nanjing, China
Search for more papers by this authorMeiping Lu
Department of Otolaryngology-Head and Neck Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
Search for more papers by this authorFeng Zheng
Huadong Medical Institute of Biotechniques, Nanjing, China
Search for more papers by this authorChangjun Wang
Huadong Medical Institute of Biotechniques, Nanjing, China
Search for more papers by this authorXiaojun Tang
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorNing Xu
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorRenjie Chen
Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China
Search for more papers by this authorDawei Zhang
Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China
Search for more papers by this authorJin Zhu
Huadong Medical Institute of Biotechniques, Nanjing, China
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorCorresponding Author
Yuan Mao
Department of Otolaryngology-Head and Neck Surgery, Jiangsu Provincial Hospital, Nanjing, China
Correspondence to: Dr. Jin Zhu, Huadong Medical Institute of Biotechniques, Nanjing 210002; The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, People's Republic of China, Tel.: 86-25-8686-3100, Fax: 86-25-8686-3100, E-mail: [email protected] or Dr. Yuan Mao, Department of Otolaryngology-Head and Neck Surgery, Jiangsu Provincial Hospital, Nanjing 210029, People's Republic of China, Tel.: 86-25-8371-2838, Fax: 86-25-8371-2838, E-mail: [email protected] or Dr. Zhenqing Feng, The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, People's Republic of China, Tel.: 86-25-8686-3100, Fax: 86-25-8686-3100, E-mail: [email protected]Search for more papers by this authorZhenqing Feng
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorHong Lin
Huadong Medical Institute of Biotechniques, Nanjing, China
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Department of Otolaryngology-Head and Neck Surgery, Jiangsu Provincial Hospital, Nanjing, China
Jiangsu Provincial Blood Center, Nanjing, China
Search for more papers by this authorHuiling Zhang
Department of Gynecology and Obstetrics, Nanjing Maternal and Children Care Hospital Affiliated to Nanjing Medical University, Nanjing, China
Search for more papers by this authorJun Wang
Department of Hematology and Oncology, Jiangsu Provincial Hospital, Nanjing, China
Search for more papers by this authorMeiping Lu
Department of Otolaryngology-Head and Neck Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China
Search for more papers by this authorFeng Zheng
Huadong Medical Institute of Biotechniques, Nanjing, China
Search for more papers by this authorChangjun Wang
Huadong Medical Institute of Biotechniques, Nanjing, China
Search for more papers by this authorXiaojun Tang
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorNing Xu
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorRenjie Chen
Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China
Search for more papers by this authorDawei Zhang
Department of Otolaryngology-Head and Neck Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China
Search for more papers by this authorJin Zhu
Huadong Medical Institute of Biotechniques, Nanjing, China
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorCorresponding Author
Yuan Mao
Department of Otolaryngology-Head and Neck Surgery, Jiangsu Provincial Hospital, Nanjing, China
Correspondence to: Dr. Jin Zhu, Huadong Medical Institute of Biotechniques, Nanjing 210002; The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, People's Republic of China, Tel.: 86-25-8686-3100, Fax: 86-25-8686-3100, E-mail: [email protected] or Dr. Yuan Mao, Department of Otolaryngology-Head and Neck Surgery, Jiangsu Provincial Hospital, Nanjing 210029, People's Republic of China, Tel.: 86-25-8371-2838, Fax: 86-25-8371-2838, E-mail: [email protected] or Dr. Zhenqing Feng, The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing 210029, People's Republic of China, Tel.: 86-25-8686-3100, Fax: 86-25-8686-3100, E-mail: [email protected]Search for more papers by this authorZhenqing Feng
The Key Laboratory of Cancer Biomarkers, Prevention & Treatment Cancer Center and The Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, Nanjing, China
Search for more papers by this authorConflict of interest: The authors declare no conflict of interest.
Abstract
Human trophoblastic cell surface antigen 2 (Trop2) has been suggested as an oncogene, which is associated with the different types of tumors. In this study, a human Fab antibody against Trop2 extracellular domain was isolated from phage library by phage display technology, and characterized by ELISA, FACS, fluorescence staining and Western blotting analysis. MTT, apoptosis assay and wound healing assay were employed to evaluate the inhibitory effects of Trop2 Fab on breast cancer cell growth in vitro, while tumor-xenograft model was employed to evaluate the inhibitory effects on breast cancer growth in vivo. The results showed that Trop2 Fab inhibited the proliferation, induced the apoptosis and suspended the migration of MDA-MB-231 cells in a dose dependent manner. The expression caspase-3 was activated, and the expression of Bcl-2 was reduced while that of Bax was elevated in MDA-MB-231 cells by treating with Trop2 Fab. In addition, Trop2 Fab inhibited the growth of breast cancer xenografts and the expression of Bcl-2 was reduced while that of Bax was elevated in xenografts. Trop2 Fab, which was isolated successfully in this research, is a promising therapeutic agent for the treatment of Trop2 expressing breast cancer.
Abstract
What's new?
Human trophoblastic cell surface antigen 2 (Trop2) is a calcium signaling molecule with oncogenic activity linked to tumor development, progression, and metastasis. Previous studies have indicated that it may be a valuable therapeutic target, as well as a prognostic or diagnostic marker. Here, a fully human Fab antibody targeting Trop2 was cloned and expressed. In breast cancer cell lines, Trop2 Fab inhibited growth, induced apoptosis, and suppressed migration, while in breast cancer xenografts, it successfully suppressed tumor growth and induced changes in the expression of cell death signaling molecules.
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