期刊
BIOORGANIC CHEMISTRY
卷 115, 期 -, 页码 -出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bioorg.2021.105266
关键词
Chromogranin; E. coli; Peptide; Colon cancer; Apoptosis; Cisplatin
资金
- Theodor Bilharz Research Institute (TBRI), Giza, Egypt [28/K]
This study successfully expressed the recombinant human CGA-N46 peptide with wide-spectrum antibacterial, fungal, and anticancer activities. The purified rhCGA-N46 peptide showed safety on normal cells and significant anticancer effects on colon cancer cells, potentially through regulating apoptotic signal pathways.
Human Chromogranin A N46 (CGA-N46) is a weak cationic alpha-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-beta-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 rhCGAN46 was safe on normal cells with IC50 = 227.74 mu g/ml (40.67 mu M) and had an obvious anticancer effect on colon cancer cells with IC50 = 1.997 mu g/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 rhCGAN46 peptide was produced successfully in the native form with promising anti-colon cancer activity.
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