Purified recombinant human Chromogranin A N46 peptide with remarkable anticancer effect on human colon cancer cells

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Okasha H.; Samir S.; Nasr S.M.

Journal: Bioorganic Chemistry

Publisher: Academic Press Inc.

Publication Date: October 2021

Volume / Issue: Volume 115

Article No.: 105266

ISSN: 452068

DOI: 10.1016/j.bioorg.2021.105266

Scopus: View on Scopus

PubMed: 34449322

Document Type: Article

Authors and Affiliations

Okasha H., Biochemistry and Molecular Biology Department, Theodor Bilharz Research Institute, Giza, Egypt; Samir S., Biochemistry and Molecular Biology Department, Theodor Bilharz Research Institute, Giza, Egypt; Nasr S.M., Biochemistry and Molecular Biology Department, Theodor Bilharz Research Institute, Giza, Egypt

Abstract

Human Chromogranin A N46 (CGA-N46) is a weak cationic α-helical peptide with wide-spectrum antibacterial, fungal, and anticancer activities. In this study, the recombinant human CGA-N46 peptide was expressed successfully in Escherichia coli. The gene of CGA-N46 was cloned into the expression vector pET-15b without a fusion tag at the N terminus and the peptide was expressed using Isopropyl-β-D-thiogalactoside (IPTG) as an inducer. Using 8 M guanidinium HCl, inclusion bodies containing the peptide were purified and solubilized. The rhCGA-N46 peptide was purified using Q-FF anion exchange column. The cytotoxicity of the purified rhCGA-N46 peptide was investigated on WI-38 human lung normal cell line. The anticancer activity was studied on human colon cancer cell line; HCT-116 cell line. Besides, the possible involvement of rhCGA-N46 peptide in regulating apoptotic signal pathways was analyzed by detecting the expression levels of BCL2, BID, and CAS-8 in the treated cells. The results concluded that the active peptide recovery was up to 41.98%. The purified rhCGA-N46 was safe on normal cells with IC50 = 227.74 µg/ml (40.67 µM) and had an obvious anticancer effect on colon cancer cells with IC50 = 1.997 µg/ml (356.6 nM). The expression level of BCL2 was down-regulated and BID and CAS-8 were up-regulated significantly in treated HCT-116 cells compared to untreated. In conclusion, the rhCGA-N46 peptide was produced successfully in the native form with promising anti-colon cancer activity. © 2021 Elsevier Inc.

Keywords

Apoptosis; Chromogranin; Cisplatin; Colon cancer; E. coli; Peptide; Cell Line; Cell Proliferation; Cell Survival; Chromogranin A; Colonic Neoplasms; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Humans; Molecular Structure; Peptides; Recombinant Proteins; Structure-Activity Relationship; antineoplastic agent; caspase 8; isopropyl thiogalactoside; protein bcl 2; protein Bid; recombinant human chromogranin A N46 peptide; unclassified drug; recombinant protein; antineoplastic activity; Article; cell inclusion; controlled study; cytotoxicity; down regulation; Escherichia coli; expression vector; HCT 116 cell line; human; human cell; IC50; molecular cloning; nonhuman; polyacrylamide gel electrophoresis; protein expression level; selectivity index; upregulation; WI-38 cell line; chemical structure; chemistry; colon tumor; dose response; drug effect; drug screening; metabolism; pathology; structure activity relation

Citation Information

Scopus Citations: 17

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