Selective Targeting of the Hedgehog Signaling Pathway by PBM Nanoparticles in Docetaxel-Resistant Prostate Cancer
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials and Reagents
2.2. Synthesis of TQ-Encapsulated PBM-NPs
2.3. Synthesis of Nanoparticle-Aptamer Conjugates
2.4. Engineering of Aptamer-Conjugated, TQ-Encapsulated PBM-NPs
2.5. Characterization of PBM-NPs
2.6. Nanoparticle Tracking Analysis (NTA) of PBM-NPs
2.7. Super Plasmon Resonance (SPR) Analysis of A10-Conjugated PBM-NPs
2.8. Immunohistochemistry
2.9. Cell Lines and Generation of Docetaxel (DTX)-Resistant Cells
2.10. MTT Assay
2.11. Immunofluorescence Evaluations
2.12. Flow Cytometry Assay
2.13. Quantitative Reverse Transcription PCR (qRT-PCR)
2.14. Western Blot Analyses
2.15. Statistical Analysis
3. Results
3.1. Characterization of the PSMA Aptamer (A10)-Conjugated, TQ-Encapsulated PBM-NPs
3.2. NMR Characterization, SPR Binding Affinity, and Cellular Uptake of TQ-Encapsulated PBM-NPs
3.3. PCa Cancer Tissues Display Altered Expressions of SHH and GLI1 that Increase with Higher Cancer Stages
3.4. PSMA Aptamer (A10)-Conjugated, TQ-Encapsulated PBM-NPs Sensitize DTX-R PCa Cells to Low Concentrations of TQ
3.5. Drug-Resistant PCa Cells Overexpress ATP-Binding Cassette (ABC) Transporter and Hh Pathway Genes
3.6. A10-Conjugated, TQ-Encapsulated PBM-NPs Downregulate PSMA and Hh Proteins in DTX-Resistant PCa Cells
3.7. A10-Conjugated, TQ-Encapsulated PBM-NPs Alter mRNA Expression of Hh Signaling Molecules in DTX-Resistant PCa Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Forward | Reverse | |
---|---|---|
18S | GGCCCTGTAATTGGAATGAGTC | CCAAGATCCAACTACGAGCTT |
ABCB1 | TGACATTTATTCAAAGTTAAAAGCA | TAGACACTTTATGCAAACATTTCAA |
PSMA | GCTTCCTCTTCGGGTGGTTT | TCCTGCTAAATGTGGTATCTGTGT |
SHH | TCT CCAGAAACTCCGAGCGA | ACTTGTCCTTACACCTCTGAGTC |
SMO | GCTACAACGTGTGCCTGGG | CATTCCGGAGGCCCGAC |
PTCH1 | TGAAATCCAAGCCCAGCGTC | CAGTAGCCTTCCCCATAGCC |
PTCH2 | CCGCCAGAGGTGATACAGAT | CCACGGTCATGGAGGTAGTC |
GLI1 | GCTGCCGTGGCCCTC | GTGTGGGGACACTCTGTCTG |
GLI2 | ACTTGATGTTCCCTGCGCTC | CTGCGGCACCAGCGT |
GLI3 | GCTCCACGACCACTGAAA AG | TGTCCAGGACTTTCATCCTCATTA |
SUFU | CCACACCTGCAAGAGAGAGT | TTGGCACTGACACCACTCAG |
KIF | GCGCCGCACTGGGGAT | CTCTGGGCCCTGCCTGG |
CK1 | TCTTCAAGTGGGCAGGGTCA | CTGCTCCGATCATCTCGTCT |
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Singh, S.K.; Gordetsky, J.B.; Bae, S.; Acosta, E.P.; Lillard, J.W., Jr.; Singh, R. Selective Targeting of the Hedgehog Signaling Pathway by PBM Nanoparticles in Docetaxel-Resistant Prostate Cancer. Cells 2020, 9, 1976. https://doi.org/10.3390/cells9091976
Singh SK, Gordetsky JB, Bae S, Acosta EP, Lillard JW Jr., Singh R. Selective Targeting of the Hedgehog Signaling Pathway by PBM Nanoparticles in Docetaxel-Resistant Prostate Cancer. Cells. 2020; 9(9):1976. https://doi.org/10.3390/cells9091976
Chicago/Turabian StyleSingh, Santosh Kumar, Jennifer B. Gordetsky, Sejong Bae, Edward P. Acosta, James W. Lillard, Jr., and Rajesh Singh. 2020. "Selective Targeting of the Hedgehog Signaling Pathway by PBM Nanoparticles in Docetaxel-Resistant Prostate Cancer" Cells 9, no. 9: 1976. https://doi.org/10.3390/cells9091976