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Updated: Sep 9, 2025
A Melanoma Patient-Derived Xenograft Model
Published on: May 20, 2019
PEPAD: A Promising Therapeutic Approach for the Treatment of Murine Melanoma (B16F10-Nex2)
Camila de Oliveira Gutierrez1, Rafael Araujo Pereira1, Claudiane Vilharroel Almeida1
1Laboratory of Protein Purification and Their Biological Functions, Faculty of Pharmaceutical Sciences, Food and Nutrition, Federal University of Mato Grosso do Sul (UFMS), Campo Grande 79070-900, MS, Brazil.
Abstract:
Background/Objectives: Cancer is one of the leading causes of death worldwide, and skin cancer is especially prevalent and lethal in Brazil. Despite advancements in treatment, there is still a need for new anticancer agents that are effective, selective, and less toxic. This study aimed to evaluate the cytotoxic and therapeutic potential of the peptide PEPAD. Methods: The cytotoxicity of PEPAD was assessed by MTT assay in murine melanoma (B16F10-Nex2), human melanoma (SK-MEL-28), breast (MCF-7), and cervical (HeLa) cancer cell lines. Selectivity was evaluated in healthy cells (RAW 264.7 and FN1). Morphological changes were analyzed by microscopy. Cell migration was assessed using scratch assays. Apoptotic features were evaluated using MitoTracker Deep Red, NucBlue, CaspACETM labeling, and flow cytometry. Immunogenic cell death was investigated by calreticulin and HMGB1 release. Molecular dynamics simulations explored peptide structure and interaction with lipid membranes. Results: PEPAD showed IC50 values of 7.4 µM and 18 µM in B16F10-Nex2 and SK-MEL-28 cells, respectively, and >60 µM in MCF-7 and HeLa cells. Low toxicity was observed in healthy cells (IC50 > 56 µM), indicating high selectivity. Apoptotic morphology and reduced cell migration were observed. Flow cytometry and fluorescence probes confirmed apoptosis and mitochondrial swelling. Calreticulin and HMGB1 release indicated immunogenic cell death. Simulations showed that PEPAD maintains a stable α-helical conformation and interacts with membranes. Conclusions: These findings highlight PEPAD's selective cytotoxicity and its potential as an anticancer agent with apoptotic and immunogenic properties, making it a promising candidate for therapeutic development.
Insights
The peptide PEPAD demonstrates selective anticancer activity against melanoma cells, inducing apoptosis and immunogenic cell death with low toxicity to healthy cells. This makes PEPAD a promising candidate for new cancer therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Cancer remains a leading global cause of mortality.
- Skin cancer poses a significant threat, particularly in Brazil.
- Novel anticancer agents with improved efficacy, selectivity, and reduced toxicity are needed.
Purpose of the Study:
- To evaluate the cytotoxic and therapeutic potential of the peptide PEPAD.
- To assess PEPAD's selectivity against cancer cell lines versus healthy cells.
- To investigate PEPAD's mechanism of action, including apoptosis and immunogenic cell death.
Main Methods:
- Cytotoxicity assessed via MTT assay in melanoma, breast, and cervical cancer cell lines.
- Selectivity evaluated in healthy cell lines.
- Apoptosis, cell migration, and immunogenic cell death markers (calreticulin, HMGB1) were analyzed.
- Molecular dynamics simulations were used to study peptide structure and membrane interactions.
Main Results:
- PEPAD exhibited selective cytotoxicity against murine (B16F10-Nex2) and human (SK-MEL-28) melanoma cells (IC50 values 7.4 µM and 18 µM, respectively).
- High selectivity was observed with low toxicity in healthy cells (IC50 > 56 µM).
- PEPAD induced apoptotic morphology, reduced cell migration, and triggered immunogenic cell death, confirmed by molecular simulations showing stable α-helical conformation and membrane interaction.
Conclusions:
- PEPAD displays selective cytotoxic effects on melanoma cells.
- The peptide possesses apoptotic and immunogenic cell death-inducing properties.
- PEPAD represents a promising candidate for the development of novel anticancer therapeutics.
