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Published on: March 6, 2019
Biochemical Characterization of a Kunitz-Type Protease Inhibitor From Mimosa regnellii and Its Effects on Melanoma
Luciana Maria Araújo Rabêlo1, Pedro Henrique de Oliveira Cardoso2,3, Leonardo Thiago Duarte Barreto Nobre4
1Laboratório De Química e Função De Proteínas Bioativas, Centro de Biociências, Departamento De Bioquímica, Universidade Federal do Rio Grande Do Norte, Natal, Rio Grande do Norte, Brazil.
Abstract:
Cancer is a term used to represent more than 100 diseases, including malignant tumors of different localizations. Among these types, cutaneous melanoma is one of the most common types of skin cancer worldwide. Newer, non-invasive, and more effective forms of treatment are urgently needed. Recently, the cancer membrane has emerged as a novel target for new anticancer drugs. Serine protease inhibitors are researched across all kingdoms; however, the Plantae kingdom is a more sustainable producer of bioactive molecules, including serine protease inhibitors. In this study, a Kunitz trypsin inhibitor (JTI) purified by RP-HPLC from Mimosa regnellii seeds was sequenced, its inhibitory activity quantified, and its inhibition constant determined. It was then tested for its ability to induce cell death via the apoptotic pathway in B16-F10 mouse melanoma cells. To evaluate its pro-apoptotic capacity, the inhibitor's effects on reactive oxygen species release, calcium release, cell morphological changes, and the inhibition of angiogenic and migratory activities of rabbit endothelial cells (RaEC) were also assessed in vitro. Tests for toxicity against normal cells were performed, confirming that JTI acts on melanoma cells with an IC50 of 0.65 µM. These results suggest that JTI has potential for use in melanoma treatment.
Insights
A plant-derived Kunitz trypsin inhibitor (JTI) shows promise for treating melanoma. This serine protease inhibitor effectively induces apoptosis in melanoma cells while sparing normal cells, suggesting potential therapeutic applications.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Cutaneous melanoma is a prevalent and aggressive skin cancer requiring novel therapeutic strategies.
- The cancer cell membrane presents a promising target for developing new anticancer drugs.
- Plant-derived serine protease inhibitors offer a sustainable source of bioactive molecules for drug discovery.
Purpose of the Study:
- To investigate the potential of a Kunitz trypsin inhibitor (JTI) from Mimosa regnellii seeds as an anti-melanoma agent.
- To characterize JTI's inhibitory activity and its mechanism of action on melanoma cells.
- To evaluate JTI's efficacy in inducing apoptosis and inhibiting angiogenesis and migration in vitro.
Main Methods:
- Purification of JTI from Mimosa regnellii seeds using RP-HPLC.
- Sequencing, activity quantification, and inhibition constant determination of JTI.
- In vitro assessment of JTI's effects on B16-F10 mouse melanoma cells, including apoptosis induction, reactive oxygen species and calcium release, and morphological changes.
- Evaluation of JTI's impact on rabbit endothelial cell (RaEC) angiogenesis and migration.
- Toxicity testing against normal cells.
Main Results:
- JTI was purified, sequenced, and characterized for its inhibitory properties.
- JTI demonstrated significant pro-apoptotic effects on B16-F10 mouse melanoma cells.
- JTI inhibited angiogenic and migratory activities of rabbit endothelial cells.
- JTI exhibited a low IC50 value of 0.65 µM against melanoma cells with minimal toxicity to normal cells.
Conclusions:
- JTI possesses potent anti-melanoma activity through the induction of apoptosis.
- JTI's ability to inhibit angiogenesis and migration further supports its therapeutic potential.
- JTI represents a promising candidate for the development of novel melanoma treatments.
