Anti-tumor effect of single-chain antibody to Reg3a in colorectal cancer

Autor: Zhiyuan Zhang, Xiaonan Wang, Tianqi Yin, Min Wang, Chen Luo, Xue-Qing Zhang, Pei Du, Lu-Ting Yu, Weihong Yu
Rok vydání: 2020
Předmět:
Models
Molecular
0301 basic medicine
Programmed cell death
Colorectal cancer
Gene Expression
Mice
Nude
Apoptosis
Pancreatitis-Associated Proteins
chemical and pharmacologic phenomena
Biology
Neogenesis
Flow cytometry
Mice
03 medical and health sciences
Antineoplastic Agents
Immunological
0302 clinical medicine
Downregulation and upregulation
Cell Movement
Peptide Library
Cell Line
Tumor
medicine
Animals
Humans
Cloning
Molecular
Cell Proliferation
Mice
Inbred BALB C
Binding Sites
medicine.diagnostic_test
Cell growth
Cell Biology
respiratory system
medicine.disease
biology.organism_classification
Recombinant Proteins
Tumor Burden
HEK293 Cells
030104 developmental biology
Cell culture
030220 oncology & carcinogenesis
Cancer research
biology.protein
Fluorouracil
Antibody
Colorectal Neoplasms
Protein Binding
Single-Chain Antibodies
Zdroj: Experimental Cell Research. 396:112278
ISSN: 0014-4827
DOI: 10.1016/j.yexcr.2020.112278
Popis: Background Regenerating protein 3a (Reg3a) is a trophic factor that functions as a stimulus in cell proliferation and neogenesis. Previous studies showed that Reg3a is ectopically upregulated in a majority of colorectal cancers (CRC) and detectable in the serum. Methods Single-chain variable fragment targeting Reg3a (scFv-Reg3a) was screened from a phage library. The bioactivity of recombinant Reg3a (rReg3a) and scFv-Reg3a were tested in LoVo and RKO cell lines using MTT, flow cytometry, wound healing and transwell analyses. Whether scFv-Reg3a inhibits tumor growth and enhances 5-fluorouracil (5-FU)-caused cell death were further examined in LoVo cell-transplanted nude BALB/c mice. Results A scFv-Reg3a from clone C2 was obtained and its binding affinity (KD) to rReg3a was determined to be 4.44 × 10-10. In cultured LoVo and RKO cells, rReg3a promoted but scFv-Reg3a inhibited cell proliferation, survival, migration and invasion. In LoVo cell-xenografted nude mice, administration of rReg3a accelerated tumor growth while scFv-Reg3a suppressed cell proliferation and reinforced 5-FU-induced cell death. Conclusion The newly developed scFv-Reg3a is an anti-cancer agent which is potent to suppress CRC cell proliferation and survival. The use of scFv-Reg3a could enhance the effectiveness of 5-FU-based chemotherapy in the cancerous treatment.
Databáze: OpenAIRE
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