In silico identiûcation and in vitro assessment of a potential anti-breast cancer activity of antimicrobial peptide retrieved from the ATMP1 Anabas testudineus ûsh peptide

A previous study has shown that synthetic antimicrobial peptides (AMPs) derived from
Anabas testudineus (ATMP1) could in-vitro inhibit the progression of breast cancer cell
lines. In this study, we are interested in studying altered versions of previous synthetic
AMPs to gain some insight into the peptides functions. The AMPs were altered and
subjected to bioinformatics prediction using four databases (ADP3, CAMP -R3, AMPfun, and
ANTICP) to select the highest anticancer activity. The bioinformatics in silico analysis led to
the selection of two AMPs, which are ATMP5 (THPPTTTTTTTTTTTYTAAPATTT) and ATMP6
(THPPTTTTTTTTTTTTTAAPARTT). The in silico analysis predicted that ATMP5 and ATMP6
have anticancer activity and lead to cell death. The ATMP5 and ATMP6 were submitted to
deep learning databases (ToxIBTL and ToxinPred2) to predict the toxicity of the peptides
and to (AllerTOP & AllergenFP) check the allergenicity. The results of databases indicated
that AMPs are non-toxic to normal human cells and allergic to human immunoglobulin. The
bioinformatics ûndings led to select the highest active peptide ATMP5, which was
synthesised and applied for in-vitro experiments using cytotoxicity assay MTT Assay,
apoptosis detection using Annexin V FTIC-A assay, and gene expression using Apoptosis
PCR Array to evaluate the AMP9s anticancer activity. The antimicrobial activity is approved
by the disc diûusion method. The in-vitro experiments analysis showed that ATMP5 had
the activity to inhibit the growth of the Breast cancer cell line (MDA-MB-231) after 48 h and
managed to arrest the cell cycle of the MDA-MB-231, apoptosis induction, and
overexpression of the p53 by interaction with the related apoptotic genes. This research
opened up new opportunities for developing potentialand selective anticancer agents
relying on antimicrobial peptides properties.