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Differences in the Inhibitory Specificity Distinguish the Efficacy of Plant Protease Inhibitors on Mouse Fibrosarcoma
Sonia Yoo Im1, Camila Ramalho Bonturi1, Adriana Miti Nakahata2
1Departamento de Bioquímica, Universidade Federal de São Paulo (UNIFESP), 04044-020 São Paulo, SP, Brazil.
Abstract:
Metastasis, the primary cause of death from malignant tumors, is facilitated by multiple protease-mediated processes. Thus, effort has been invested in the development of protease inhibitors to prevent metastasis. Here, we investigated the effects of protease inhibitors including the recombinant inhibitors rBbKI (serine protease inhibitor) and rBbCI (serine and cysteine inhibitor) derived from native inhibitors identified in Bauhinia bauhinioides seeds, and EcTI (serine and metalloprotease inhibitor) isolated from the seeds of Enterolobium contortisiliquum on the mouse fibrosarcoma model (lineage L929). rBbKI inhibited 80% of cell viability of L929 cells after 48 h, while EcTI showed similar efficacy after 72 h. Both inhibitors acted in a dose and time-dependent manner. Conversely, rBbCI did not significantly affect the viability of L929 cells. Confocal microscopy revealed the binding of rBbKI and EcTI to the L929 cell surface. rBbKI inhibited approximately 63% of L929 adhesion to fibronectin, in contrast with EcTI and rBbCI, which did not significantly interfere with adhesion. None of the inhibitors interfered with the L929 cell cycle phases. The synthetic peptide RPGLPVRFESPL-NH2, based on the BbKI reactive site, inhibited 45% of the cellular viability of L929, becoming a promising protease inhibitor due to its ease of synthesis.
Insights
Protease inhibitors rBbKI and EcTI effectively reduced fibrosarcoma cell viability and adhesion. A synthetic peptide based on rBbKI also showed promise as a novel protease inhibitor.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Metastasis, a major cause of cancer mortality, involves protease-mediated processes.
- Protease inhibitors are developed to combat cancer metastasis.
- Native inhibitors from *Bauhinia bauhinioides* and *Enterolobium contortisiliquum* seeds offer potential therapeutic leads.
Purpose of the Study:
- To investigate the anti-metastatic potential of recombinant protease inhibitors rBbKI, rBbCI, and EcTI.
- To evaluate the effects of these inhibitors on L929 fibrosarcoma cells.
- To explore the therapeutic promise of a synthetic peptide derived from rBbKI.
Main Methods:
- Treatment of L929 fibrosarcoma cells with rBbKI, rBbCI, and EcTI.
- Assessment of cell viability, adhesion to fibronectin, and cell cycle progression.
- Confocal microscopy for inhibitor-cell surface binding analysis.
- Synthesis and testing of a peptide inhibitor based on the rBbKI reactive site.
Main Results:
- rBbKI and EcTI significantly inhibited L929 cell viability in a dose- and time-dependent manner.
- rBbKI demonstrated significant inhibition of L929 cell adhesion to fibronectin.
- rBbCI showed no significant effect on cell viability or adhesion.
- The synthetic peptide RPGLPVRFESPL-NH2 inhibited L929 cell viability by 45%.
Conclusions:
- Recombinant inhibitors rBbKI and EcTI exhibit anti-cancer properties against fibrosarcoma.
- rBbKI is particularly effective in reducing cell viability and adhesion.
- A synthetic peptide inhibitor offers a promising, easily synthesized alternative for cancer therapy.
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