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Hybrid Anticancer Peptides DN1 and DN4 Exert Selective Cytotoxicity Against Hepatocellular Carcinoma Cells by Inducing Both Intrinsic and Extrinsic Apoptotic Pathways

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Abstract

Bioactive peptides have emerged as promising therapeutic alternatives in pharmaceutical industry, especially to fight cancer. Here we aim to investigate the in vitro anti-proliferative activities and cell death mechanism of two series of hybrid peptides designed based on IsCT peptide (from Opithacantus madagascariensis) and A4 peptide (from Escherichia coli bacterial membrane anchor and aurein 1.2) against hepatocellular carcinoma cells. The ND series of hybrid peptides (IC50 ranging from 22 to 72 µg/ml) generally displayed higher cytotoxicity on all cell lines tested as compared to the DN peptides (IC50 ≥ 105 µg/ml). Among these peptides, DN1 and DN4 exhibited observable selectivity against HepG2 cells in a dose-dependent manner in the absence of cytostatic activity. DN1 and DN4 were shown to effectively kill HepG2 cells via minimum up-regulation of p53 accompanied with concurrent activation of both intrinsic and extrinsic apoptosis pathways that were cross-linked by Bid protein. We show that DN1 and DN4 induce selective cytotoxicity against HepG2 cells by triggering activation of intrinsic and extrinsic apoptosis pathways. Such concomitant activation of both apoptosis pathways to achieve efficient cytotoxicity may also reduce the likelihood of drug resistance development. In addition, cytotoxicity and selectivity of the hybrid peptides against HepG2 cells appeared to correlate with peptides’ cationicity and hydrophobicity. DN1 and DN4 showed potential to be further explored as an alternative in hepatocellular carcinoma and other primary cancer treatment. Further modulation of physicochemical properties of DN1 and DN4 can be done in search of novel ACP variants with enhanced potency as anticancer therapeutics.

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The datasets used and/or analyzed in the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We acknowledge the support provided by Taylor’s University Centralised Laboratories throughout the research studies.

Funding

This study was supported by Taylor’s University, Lakeside Campus, Emerging Research Grant (TRGS/ERFS/2/2018/SBS/013) and Fundamental Research Grant Scheme (FRGS/1/2018/STG05/UNIM/03/1), Ministry of Education, Malaysia.

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  1. School of Biosciences, Faculty of Health and Medical Sciences, Taylor’s University, Lakeside Campus, Subang Jaya, Selangor, Malaysia

    Chu Xin Ng, Yin Sim Tor & Sau Har Lee

  2. School of Biosciences, Faculty of Science and Engineering, University of Nottingham Malaysia, Semenyih, Selangor, Malaysia

    Cheng Foh Le

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CXN carried out all experiments, participated in all the statistical analysis and drafted the manuscript. CFL and YST participated in the study conceptualization, reviewed, and revised the manuscript. SHL conceived and administered the project design, supervised, reviewed, and revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Sau Har Lee.

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Ng, C.X., Le, C.F., Tor, Y.S. et al. Hybrid Anticancer Peptides DN1 and DN4 Exert Selective Cytotoxicity Against Hepatocellular Carcinoma Cells by Inducing Both Intrinsic and Extrinsic Apoptotic Pathways. Int J Pept Res Ther 27, 2757–2775 (2021). https://doi.org/10.1007/s10989-021-10288-8

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