Pseudomonas Exotoxin A Based Toxins Targeting Epidermal Growth Factor Receptor for the Treatment of Prostate Cancer
Abstract
:1. Introduction
2. Results
2.1. Cloning, Expression and Purification of EGF and the Targeted Toxins EGF-PE40 and EGF-PE24mut
2.2. Binding of EGF-PE40 and EGF-PE24mut to Different PCa Cell Lines
2.3. EGF-PE40 and EGF-PE24mut Inhibit Protein Biosynthesis
2.4. EGF-PE40 and EGF-PE24mut Reduce Cell Viability and Induce Apoptosis
3. Discussion
4. Materials and Methods
4.1. Cells and Chemicals
4.2. Cloning, Expression and Purification of EGF and the Anti-EGFR Immunotoxins
4.3. Preparation of Cell Lysates
4.4. SDS-PAGE and Western Blot
4.5. Flow Cytometry
4.6. Inhibition of Protein Biosynthesis
4.7. WST-1 Cell Viability Assay
4.8. Bright-Field Microscopy
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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LNCaP | DU145 | PC-3 | CHO | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
IC50(nM) | IC50(nM) | IC50(nM) | IC50(nM) | |||||||||
24h | 48h | 72h | 24h | 48h | 72h | 24h | 48h | 72h | 24h | 48h | 72h | |
EGF-PE40 | 2.31 | 0.04 | 0.008 | 2.71 | 0.31 | 0.18 | 9.67 | 0.64 | 0.86 | >14.5 | >14.5 | >14.5 |
EGF-PE24mut | >20.6 | 1.91 | 0.97 | >20.6 | 8.52 | 5.27 | >20.6 | 8.80 | 9.59 | >20.6 | >20.6 | >20.6 |
Erlotinib | nd | >5750 | nd | nd | nd | >5750 | nd | nd | >5750 | nd | nd | >5750 |
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Fischer, A.; Wolf, I.; Fuchs, H.; Masilamani, A.P.; Wolf, P. Pseudomonas Exotoxin A Based Toxins Targeting Epidermal Growth Factor Receptor for the Treatment of Prostate Cancer. Toxins 2020, 12, 753. https://doi.org/10.3390/toxins12120753
Fischer A, Wolf I, Fuchs H, Masilamani AP, Wolf P. Pseudomonas Exotoxin A Based Toxins Targeting Epidermal Growth Factor Receptor for the Treatment of Prostate Cancer. Toxins. 2020; 12(12):753. https://doi.org/10.3390/toxins12120753
Chicago/Turabian StyleFischer, Alexandra, Isis Wolf, Hendrik Fuchs, Anie Priscilla Masilamani, and Philipp Wolf. 2020. "Pseudomonas Exotoxin A Based Toxins Targeting Epidermal Growth Factor Receptor for the Treatment of Prostate Cancer" Toxins 12, no. 12: 753. https://doi.org/10.3390/toxins12120753