Repurposing an antimicrobial arginine-rich decapeptide as a novel anticancer agent: Evidence from in vitro and in vivo breast cancer models

Document Type : Original Article

Authors

Department of Biology, Yazd University, Yazd, Iran

Abstract

Objective(s): Arginine-rich peptides have attracted interest because of their ability to interact with negatively charged cancer cell membranes. The present study aimed to evaluate the anticancer properties of a novel antimicrobial arginine-rich decapeptide (RL10) in vitro and in vivo. 
Materials and Methods: Cytotoxicity was assessed in breast (4T1), colon (SW480), and normal fibroblast (NIH3T3) cell lines by MTT assay following 48- and 72-hr treatment (6-400 µg/ml). Membrane integrity was examined in 4T1 cells after 48 hr using lactate dehydrogenase (LDH) release. Flow cytometry was applied to determine apoptosis induction and alterations in cell cycle distribution. Caspase 3/7 activity was also evaluated. Apoptosis-related gene expressions were analyzed using RT-PCR. The antitumor effect of RL10 was assessed in a murine 4T1 model, followed by histopathological analysis using H&E staining.
Results: RL10 significantly declined cancer cell viability in a dose-dependent manner, with no toxicity on normal cells. No increase in LDH release was detected. Flow cytometry revealed 28.9% apoptosis induction and cell cycle arrest at the S and G2/M phases. Marked upregulation of the Casp9 gene suggested possible activation of the intrinsic apoptotic pathway; however, Bax, Bcl2, p53, and Casp8 expressions remained unchanged. In vivo, two weeks’ treatment with the peptide suppressed tumor growth by nearly 39% (P˂0.01), accompanied by increased apoptotic figures and reduced mitotic counts. 
Conclusion: These findings provide the first evidence that an arginine-rich short peptide exhibits anticancer activity through apoptosis induction and tumor growth inhibition, underscoring its potential as a candidate for cancer therapy or drug delivery systems.

Keywords

Main Subjects

References

1. Wilkinson L. Understanding breast cancer as a global health concern. Br J Radiol 2022; 95: 1-8. 
2. Sood A, Jothiswaran V V, Singh A, Sharma A. Anticancer peptides as novel immunomodulatory therapeutic candidates for cancer treatment. Explor Target Anti-tumor Ther 2024; 5: 1074-1099. 
3. Bauso LV, La Fauci V, Munaò S, Bonfiglio D, Armeli A, Maimone N, et al. Biological activity of natural and synthetic peptides as anticancer agents. Int J Mol Sci 2024; 25: 1-19. 
4. Sarabi N, Chamani R, Assareh E, Saberi O, Asghari SM. Combination therapy in cancer: Doxorubicin in combination with an N-terminal peptide of endostatin suppresses angiogenesis and stimulates apoptosis in the breast cancer. Int J Mol Cell Med 2023; 12: 1-15 
5. Chamani R, Saberi O, Fathinejad F. An arresten-derived anti-angiogenic peptide triggers apoptotic cell death in endothelial cells. Mol Biol Rep 2024; 51: 1-11
6. Nhàn NTT, Yamada T, Yamada KH. Peptide-based agents for cancer treatment: Current applications and future directions. Int J Mol Sci 2023; 24: 1-15. 
7. Meloni BP, Mastaglia FL, Knuckey NW. Cationic arginine-rich peptides (CARPs): A novel class of neuroprotective agents with a multimodal mechanism of action. Front Neurol 2020; 11: 1-20 
8. Liscano Y, Oñate-Garzón J, Delgado JP. Peptides with dual antimicrobial–anticancer activity: Strategies to overcome peptide limitations and rational design of anticancer peptides. Molecules 2020; 25: 1-20 
9. Bucataru C, Ciobanasu C. Antimicrobial peptides: Opportunities and challenges in overcoming resistance. Microbiol Res 2024; 286: 1-15. 
10. Varela-Quitián YF, Mendez-Rivera FE, Bernal-Estévez DA. Cationic antimicrobial peptides: Potential templates for anticancer agents. Front Med 2025; 12: 1-16. 
11. Dong Z, Zhang X, Zhang Q, Tangthianchaichana J, Guo M, Du S, et al. Anticancer mechanisms and potential anticancer applications of antimicrobial peptides and their nano agents. Int J Nanomedicine 2024; 19: 1017-1039. 
12. Agrillo B, Porritiello A, Gratino L, Balestrieri M, Proroga YT, Mancusi A, et al. Antimicrobial activity, membrane interaction and structural features of short arginine-rich antimicrobial peptides. Front Microbiol 2023; 14: 1-20. 
13. Chamani R, Taleqani MH, Imanpour A, and Khatami M. New insights into short peptides derived from the collagen NC1 α1, α2, and α3 (IV) domains: An experimental and MD simulations study. Biochim Biophys Acta - Proteins Proteomics 2022; 1870: 1-10. 
14. Hu C, Chen X, Huang Y, Chen Y. Synergistic effect of the pro-apoptosis peptide kla-TAT and the cationic anticancer peptide HPRP-A1. Apoptosis 2018; 23: 132-142. 
15. Dai Y, Cai X, Shi W, Bi X, Su X, Pan M, et al. Pro-apoptotic cationic host defense peptides rich in lysine or arginine to reverse drug resistance by disrupting tumor cell membrane. Amino Acids 2017; 49: 1601-1610. 
16. Chamani R, Darvand-Araghi MH. Disulfide bond : A critical element in the structure and function of a collagen IV-derived antiangiogenic peptide. 2025; 5: 289-303. 
17. Agrillo B, Proroga Y, Gogliettino M, Balestrieri M, Tate R, Nicolais L, et al. A safe and multitasking antimicrobial decapeptide: The road from de novo design to structural and functional characterization. Int J Mol Sci 2020; 21: 1-22. 
18. Van Tonder A, Joubert AM, Cromarty AD. Limitations of the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay when compared to three commonly used cell enumeration assays. BMC Res Notes 2015; 8: 1-10. 
19. Pan W R, Chen P W, Chen Y L, Hsu H C, Lin C C, Chen W J. Bovine lactoferricin B induces apoptosis of human gastric cancer cell line AGS by inhibition of autophagy at a late stage. J Dairy Sci 2013; 96: 7511-7520. 
20. Abdel-Salam M, Pinto B, Cassali G, Bueno L, Pêgas G, Oliveira F, et al. LyeTx Ib peptide attenuates tumor burden and metastasis in a mouse 4T1 breast cancer model. Antibiotics 2021; 10: 1-28. 
21. Theansungnoen T, Maijaroen S, Jangpromma N, Yaraksa N, Daduang S, Temsiripong T, et al. Cationic antimicrobial peptides derived from Crocodylus siamensis leukocyte extract, revealing anticancer activity and apoptotic induction on human cervical cancer cells. Protein J 2016; 35: 202-211. 
22. Hilchie AL, Sharon AJ, Haney EF, Hoskin DW, Bally MB, Franco OL, et al. Mastoparan is a membranolytic anti-cancer peptide that works synergistically with gemcitabine in a mouse model of mammary carcinoma. Biochim Biophys Acta (BBA)-Biomembranes 2016; 1858: 3195-3204. 
23. Hadianamrei R, Tomeh MA, Brown S, Wang J, Zhao X. Rationally designed short cationic α-helical peptides with selective anticancer activity. J Colloid Interface Sci 2022; 607: 488-501. 
24. Arias M, Haney EF, Hilchie AL, Corcoran JA, Hyndman ME, Hancock REW, et al. Selective anticancer activity of synthetic peptides derived from the host defence peptide tritrpticin. Biochim Biophys Acta (BBA)-Biomembranes 2020; 1862: 1-10. 
25. Nguyen T, Guo R, Chai J, Wu J, Liu J, Chen X, et al. Smp24, a scorpion-venom peptide, exhibits potent antitumor effects against hepatoma HepG2 cells via multi-mechanisms in vivo and in vitro. Toxins (Basel) 2022; 14: 1-19. 
26. Singhal M, Shaha S, Katsikogianni M. Comparative analysis of cytotoxicity assays, from traditional to modern approaches. From edited volume: Cytotoxicity-a crucial toxicity test for in vitro experiments (Edited by: Erkekoglu P.). IntechOpen 2025. p. 1-54 
27. Wang H, Guo M, Wei H, Chen Y. Targeting p53 pathways: Mechanisms, structures, and advances in therapy. Signal Transduct Target Ther 2023; 8: 1-35. 
28. D’costa M, Bothe A, Das S, Kumar SU, Gnanasambandan R, Doss CGP. CDK regulators: Cell cycle progression or apoptosis-Scenarios in normal cells and cancerous cells. Adv Protein Chem Struct Biol 2023; 135: 125-177. 
29. Yue J, López JM. Understanding MAPK signaling pathways in apoptosis. Int J Mol Sci 2020; 21: 1-22. 
30. Pungsrinont T, Kallenbach J, Baniahmad A. Role of PI3K-AKT-mTOR pathway as a pro-survival signaling and resistance-mediating mechanism to therapy of prostate cancer. Int J Mol Sci 2021; 22: 1-25. 
31. Yu Y, Feng X-H. TGF-β signaling in cell fate control and cancer. Curr Opin Cell Biol 2019; 61: 56-63. 
32. Dong L, Li Y, Zhang Y, Su S. Cationic antimicrobial peptide CC34 potential anticancer and apoptotic induction on cancer cells. Amino Acids 2025; 57: 1-13. 
33. Mustafa M, Ahmad R, Tantry IQ, Ahmad W, Siddiqui S, Alam M, et al. Apoptosis: A comprehensive overview of signaling pathways, morphological changes, and physiological significance and therapeutic implications. Cells 2024; 13: 1-20.
34. Liu S, Aweya JJ, Zheng L, Zheng Z, Huang H, Wang F, et al. LvHemB1, a novel cationic antimicrobial peptide derived from the hemocyanin of Litopenaeus vannamei, induces cancer cell death by targeting mitochondrial voltage-dependent anion channel 1. Cell Biol Toxicol 2022; 38: 87-110. 
Iranian Journal of Basic Medical Sciences
Volume 29, Issue 5
May 2026
Pages 736-743

Files

Share

How to cite

Statistics