Abstract
Liver cancer is a worldwide issue that also affects the Malaysian population. The occurrence is closely related to risk factors like chronic infections and environmental exposures. Due to the toxicity of conventional therapeutic drugs for liver cancer, bioactive peptides have emerged as a popular alternative anticancer agent. Although the full-length pardaxin from Pardachirus marmoratus was proven with anticancer effects, its concurrent haemolytic effects are yet to be resolved. Therefore, this study utilized in silico and in vitro analyses to assess cytotoxic effects induced by the shortened pardaxin derivatives. The in silico findings led to the discovery of a series of shortened pardaxin derivatives with 13 amino acids, where single residue replacement prediction by bioinformatics tools was done on the shortened sequences. Among the top five shortened derivatives, the derivative where amino acid threonine was replaced by proline, was identified as the most potential candidate, namely LL13. The LL13 peptide was predicted with improved anticancer effects, non-toxic, and alleviated haemolytic effects as compared to its parental peptide. The subsequent cytotoxicity testing further validated its selective toxicity against liver cancer cells, HepG2 cells, with relatively lower killing effects on the normal cells, Vero cells. These in vitro findings validated the in silico predictions and also indicated that this peptide has potential as an anticancer drug with selective targeting capabilities. In conclusion, this study has highlighted the potential of using a combination of in silico and in vitro approaches to discover potentially shortened peptides as a novel therapeutic option for liver cancer treatment.
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Abbreviations
- NAFLD:
-
Non-alcoholic fatty liver disease
- HCC:
-
Hepatocellular carcinoma
- CCA:
-
Cholangiocarcinoma
- SVM:
-
Support vector machine
- CPP:
-
Cell-penetrating peptide
- PDB:
-
Protein Data Bank
- ATCC:
-
American Type Culture Collection
- DMEM:
-
Dulbecco's Modified Eagle's Medium
- FBS:
-
Fetal bovine serum
- MTT:
-
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide
- DMSO:
-
Dimethyl sulfoxide
- IC50 :
-
Half maximal inhibitory concentration
- SEM:
-
Standard error of mean
- AntiCP:
-
Anticancer peptide
- FASL:
-
Fas ligand
- FAS-FADD:
-
Fas-associated protein with death domain
- ROS:
-
Reactive oxygen species
- UPR:
-
Unfolded protein response
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Acknowledgements
This work was supported by Taylor’s University through its TAYLOR’S RESEARCH SCHOLARSHIP Programme and National Cancer Council Malaysia (MAKNA) Cancer Research Collaboration Fund 2022 (MAKNA/2022/SBS/001).
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KMW and YHW has contributed equally to this study. KMW conducted in vitro validation assays. YHW carried out the in silico predictions. KMW and YHW drafted and edited the manuscript. SHL conceived, reviewed and edited the manuscript. All authors read and approved the final manuscript.
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Wong, K.M., Wong, Y.H. & Lee, S.H. In Silico Discovery of LL13, a Shortened Pardaxin 6 Peptide Derivative with Anti-proliferative Activity. Int J Pept Res Ther 30, 38 (2024). https://doi.org/10.1007/s10989-024-10615-9
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DOI: https://doi.org/10.1007/s10989-024-10615-9