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Cloning, expression of the truncation of recombinant peroxidase derived from millet bran and its reversal effects on 5-Fu resistance in colorectal cancer.
- Source :
-
International journal of biological macromolecules [Int J Biol Macromol] 2019 Jul 01; Vol. 132, pp. 871-879. Date of Electronic Publication: 2019 Mar 19. - Publication Year :
- 2019
-
Abstract
- A novel peroxidase (FMBP) was extracted and purified from foxtail millet bran in our previous study and it possessed excellent anti-colon cancer activity both in vitro and in vivo. However, the active fragment of FMBP responsible for the anti-colon cancer effects remains unclear. In present, three different truncated sequences of FMBP were designed and cloned into a plasmid vector (pMal-s). Three recombinant segments were successfully expressed in host strain Escherichia coli DH5α induced by IPTG (0.3 mM) at 37 °C for 4 h, respectively named MBP-FMBP-1, MBP-FMBP-2 and MBP-FMBP-3. MTT assay showed that only MBP-FMBP-2 possessed significant anti-colon cancer activity, and its anti-colon cancer activity was equivalent to FMBP. Further, the results showed that MBP-FMBP-2 significantly reversed the 5-Fu resistance in human colorectal cancer HCT-8/Fu cell through inhibiting cell proliferation, promoting cell apoptosis and increasing the intracellular accumulation of 5-Fu. RT-PCR and western blot assays showed that MBP-FMBP-2 also decreased the expression levels of multi-drug resistance protein 1 (MRP1), P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP). These results indicate that MBP-FMBP-2 is the effective segment of FMBP which exhibits anti-colon cancer activity and has potential as an outstanding chemotherapeutic agent against colon cancer.<br /> (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Subjects :
- Apoptosis drug effects
Cell Line, Tumor
Cell Proliferation drug effects
Cloning, Molecular
Drug Resistance, Multiple drug effects
Gene Expression
Gene Expression Regulation, Neoplastic drug effects
Genetic Engineering
Humans
Peroxidase chemistry
Protein Domains
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins pharmacology
Colorectal Neoplasms pathology
Drug Resistance, Neoplasm drug effects
Fluorouracil pharmacology
Millets chemistry
Peroxidase genetics
Peroxidase pharmacology
Subjects
Details
- Language :
- English
- ISSN :
- 1879-0003
- Volume :
- 132
- Database :
- MEDLINE
- Journal :
- International journal of biological macromolecules
- Publication Type :
- Academic Journal
- Accession number :
- 30902718
- Full Text :
- https://doi.org/10.1016/j.ijbiomac.2019.03.111