Abstract
This study demonstrates the anticancer potential of RF13 peptide derived from vacuolar protein sorting associated protein 26B (VSP26B), which was obtained from transcriptome data of freshwater teleost, striped murrel (Channa striatus). In total, 28 different peptide segments were randomly predicted from VSP26B and their anticancer property was predicted through mACPpred algorithm. Based on the scoring value, out of 28, 3 peptides namely SA11, RF13 and DI14 were shortlisted for the primary physicochemical property screening. Also, the shortlisted peptides were validated for anticancer protein receptors using molecular docking. Based on the overall bioinformatics analysis, RF13 ((1)RRGKGGRRVTMSF(13)) peptide was selected to further investigate the anticancer potential at the molecular biology lab. In vitro experiment showed that RF13 inhibit the proliferation of human laryngeal epithelial cell (HEp-2). Further, microscopic observations confirmed the influence of peptides on the cell morphology that showed apoptosis due to Hoechst 33342 staining, which altogether demonstrated the anticancer effect of the examined peptide, RF13. The molecular mechanism of the peptide's anticancer property was analyzed over cell cycle analysis. It showed that the peptide arrested the SubG1 and G1 phase of Hep-2 cells in a dose-dependent manner. The qPCR assay exhibited the apoptotic genes, including caspase-3 and caspase-9 expression significantly (p < 0.05) in an unregulated manner. Based on the obtained results, we propose that RF13 is potential enough as an anticancer agent; however, further direction needs to be focused on the possible specific target mechanism and its therapeutic approach towards anticancer drug development.