Back to Search
Start Over
Single-chain Antibody Against Reg4 Suppresses Gastric Cancer Cell Growth and Enhances 5-FU-induced Cell Death in vitro
- Source :
- Anti-Cancer Agents in Medicinal Chemistry. 19:610-619
- Publication Year :
- 2019
- Publisher :
- Bentham Science Publishers Ltd., 2019.
-
Abstract
- Background:Regenerating islet-derived gene family member 4 (Reg4), a well-investigated growth factor in the regenerative pancreas, has recently been reported to be highly associated with a majority of gastrointestinal cancers. Pathological hyper-expression or artificial over-expression of Reg4 causes acceleration of tumor growth, migration, and resistance to chemotherapeutic 5-Fluorouracil (5-FU). Until now, no method has been successfully established for eliminating the effects of Reg4 protein.Methods:This study reports the production of an engineered immunoglobin, a single-chain variable fragment (scFv-Reg4), to specifically bind Reg4 and block the bioactivity. The complementary-determining regions (CDRs) against Reg4 were assigned using MOE and ZDOCK servers. The binding affinity (KD) was determined by bio-layer interferometry (BLI). MKN45 and AGS cell proliferation was determined by Thiazolyl blue tetrazolium bromide (MTT) method and the cell apoptosis was detected by flow cytometry assay.Results:The KD of scFv-Reg4 to Reg4 was determined to be 1.91×10-8. In MKN45 and AGS cell lines, scFv- Reg4 depressed Reg4-stimulated cell proliferation and the inhibitory rates were 27.7±1.5% and 17.3±2.6%, respectively. Furthermore, scFv significantly enhanced 5-FU-induced cell death, from 23.0±1.0% to 28.4±1.2% in MKN45 and 28.2±0.7% to 36.6±0.6% in AGS cells. Treatment with scFv alone could lyse cancer cells to a certain extent, but no significance has been observed.Conclusion:The single-chain antibody (scFv-Reg4) significantly inhibited gastric cancer cell proliferation and synergistically enhanced the lethal effect of 5-FU. Thus, traditional chemo-/radio- therapeutics supplemented with scFv-Reg4 may provide advances in the strategy for gastrointestinal cancer treatment.
- Subjects :
- Antimetabolites, Antineoplastic
Cancer Research
Programmed cell death
medicine.medical_treatment
Pancreatitis-Associated Proteins
Flow cytometry
03 medical and health sciences
0302 clinical medicine
Stomach Neoplasms
Cell Line, Tumor
medicine
Humans
Cell Proliferation
030304 developmental biology
Pharmacology
0303 health sciences
Cell Death
biology
medicine.diagnostic_test
Cell growth
Chemistry
Growth factor
Cell culture
Apoptosis
030220 oncology & carcinogenesis
Cancer cell
biology.protein
Cancer research
Molecular Medicine
Fluorouracil
Antibody
Single-Chain Antibodies
Subjects
Details
- ISSN :
- 18715206
- Volume :
- 19
- Database :
- OpenAIRE
- Journal :
- Anti-Cancer Agents in Medicinal Chemistry
- Accession number :
- edsair.doi.dedup.....b29b2e565b76aa8ae46f53957fce1a44