Methyl Jasmonate Effect on Betulinic Acid Content and Biological Properties of Extract from Senna obtusifolia Transgenic Hairy Roots
Abstract
:1. Introduction
2. Results
2.1. Transgenic Hairy Root Growth Index
2.2. Effect of Elicitation on Protein Profile of Senna Obtusifolia Transgenic Hairy Roots
2.3. HPLC Analysis
2.4. Effect of Transgenic Hairy Roots of Senna obtusifolia Extracts on U87MG, DU-145 and A549 Cancer Cells Lines
2.5. Apoptosis and Necrosis Determination Using Flow Cytometry
2.6. Loss of Mitochondrial Membrane Potential (MMP)
2.7. Chromosomal DNA Fragmentation
2.8. DNA Damage Measured by Comet Assay
2.9. Influence on the Caspase-3/7 Activity
2.10. Topoisomerase I Inhibition Assay with Hairy Root Extract
3. Discussion
4. Materials and Methods
4.1. Establishment of Hairy Root Cultures
4.2. Optimization of Hairy Root Culture Conditions in the Bioreactor
4.3. Elicitor Treatment
4.4. Hairy Roots Growth Index Determination
4.5. Hairy Root Protein Isolation
4.6. SDS-PAGE Protein Electrophoresis
4.7. Plant Extract Preparation
4.8. HPLC Determination of Betulinic Acid
4.9. Cell Cultures
4.10. Cell Viability
4.11. Apoptosis/Necrosis Detection by Flow Cytometry
4.12. Mitochondrial Membrane Potential (MMP)
4.13. Chromosomal DNA Fragmentation
4.14. DNA Damage Measured by Comet Assay
4.15. Effect on the Activity of Caspase-3/7
4.16. Topoisomerase I Inhibition Assay with Hairy Root Extract
4.17. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Kowalczyk, T.; Sitarek, P.; Merecz-Sadowska, A.; Szyposzyńska, M.; Spławska, A.; Gorniak, L.; Bijak, M.; Śliwiński, T. Methyl Jasmonate Effect on Betulinic Acid Content and Biological Properties of Extract from Senna obtusifolia Transgenic Hairy Roots. Molecules 2021, 26, 6208. https://doi.org/10.3390/molecules26206208
Kowalczyk T, Sitarek P, Merecz-Sadowska A, Szyposzyńska M, Spławska A, Gorniak L, Bijak M, Śliwiński T. Methyl Jasmonate Effect on Betulinic Acid Content and Biological Properties of Extract from Senna obtusifolia Transgenic Hairy Roots. Molecules. 2021; 26(20):6208. https://doi.org/10.3390/molecules26206208
Chicago/Turabian StyleKowalczyk, Tomasz, Przemysław Sitarek, Anna Merecz-Sadowska, Monika Szyposzyńska, Aleksandra Spławska, Leslaw Gorniak, Michał Bijak, and Tomasz Śliwiński. 2021. "Methyl Jasmonate Effect on Betulinic Acid Content and Biological Properties of Extract from Senna obtusifolia Transgenic Hairy Roots" Molecules 26, no. 20: 6208. https://doi.org/10.3390/molecules26206208