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Preparation of a novel EGFR specific immunotoxin and its efficacy of anti-colorectal cancer in vitro and in vivo.

Authors
  • Wu, Shuifa1, 2
  • Deng, Cuimin1, 3
  • Zhang, Caiyun1
  • Xiong, Jiani4, 5
  • Gu, Xiaofan1
  • Wang, Ze1
  • Tu, Jingjing1
  • Xie, Jieming6
  • 1 Department of Pharmacology, College of Pharmacy, Fujian Medical University, Fuzhou, Fujian Province, People's Republic of China. , (China)
  • 2 Department of Pharmacy, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou, Fujian Province, People's Republic of China. , (China)
  • 3 Department of Pharmacy, QuanZhou Women's and Children's Hospital, Quanzhou, Fujian Province, People's Republic of China. , (China)
  • 4 Department of Medical Oncology, Fujian Cancer Hospital and Fujian Medical University Cancer Hospital, Fuzhou, Fujian Province, People's Republic of China. , (China)
  • 5 Cancer Bio-Immunotherapy Center, Fujian Cancer Hospital and Fujian Medical University Cancer Hospital, Fuzhou, Fujian Province, People's Republic of China. , (China)
  • 6 Department of Pharmacology, College of Pharmacy, Fujian Medical University, Fuzhou, Fujian Province, People's Republic of China. [email protected] , (China)
Type
Published Article
Journal
Clinical & Translational Oncology
Publisher
Springer-Verlag
Publication Date
Aug 01, 2021
Volume
23
Issue
8
Pages
1549–1560
Identifiers
DOI: 10.1007/s12094-020-02548-8
PMID: 33474678
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Epithelial growth factor receptor (EGFR), as a malignancy marker, is overly expressed in multiple solid tumors including colorectal neoplasms, one of the most prevalent malignancies worldwide. The main objective of this study is to enhance the efficacy of anti-tumor therapy targeting EGFR by constructing a novel EGFR-specific immunotoxin (C-CUS245C) based on Cetuximab and recombinant Cucurmosin (CUS245C). E. coli BL21 (DE3) PlysS (E. coli) was used to express CUS245C with a cysteine residue inserting to the C-terminus of Cucurmosin. Then immobilized metal ion affinity chromatography (IMAC) was used to purify CUS245C. The chemical conjugation method was used for the preparation of C-CUS245C. Then dialysis and IMAC were used to purify C-CUS245C. Western blot as well as SDS-PAGE was carried out to characterize the formation of C-CUS245C. At last the anti-colorectal cancer activity of C-CUS245C was investigated in vitro and in vivo. CUS245C with high purity could be obtained from the prokaryotic system. C-CUS245C was successfully constructed and highly purified. The cytotoxicity assays in vitro showed a significant proliferation inhibition of C-CUS245C on EGFR-positive cells for 120 h with IC50 values less than 0.1 pM. Besides, the anti-tumor efficacy of C-CUS245C was remarkably more potent than that of Cetuximab, CUS245C, and C + CUS245C (P < 0.001). Whereas the cytotoxicity of C-CUS245C could hardly be detected on EGFR-null cell line. Our results also showed that C-CUS245C had efficacy of anti-colorectal cancer in mouse xenograft model, indicating the therapeutic potential of C-CUS245C for the targeted therapy of colorectal neoplasms. C-CUS245C exhibits potent and EGFR-specific cytotoxicity. Insertional mutagenesis technique is worthy to be adopted in the preparation of immunotoxin. Immunotoxin can be highly purified through dialysis followed by IMAC.

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