A Comprehensive Analysis of FUT8 Overexpressing Prostate Cancer Cells Reveals the Role of EGFR in Castration Resistance
Abstract
:1. Introduction
2. Results
2.1. Characterization of FUT8 Expressing Prostate Cancer Cells using LC MS/MS Mass Spectrometry
2.2. Cell Surface Receptors and FUT8 Overexpression
2.3. Overexpression of FUT8 Suppress the PSA Production and Increase Drug Resistance in Prostate Cancer Cells
2.4. Overexpression of FUT8 Promotes DNA Replication and Cell Cycle Progression
2.5. Regulation of EGFR Expression by FUT8
2.6. Castration-Induced FUT8 Overexpression Correlates with EGFR Expression in Xenograft Model
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Reagents
4.2. Western Blot Analysis
4.3. Protein and Peptide Extraction from Cells for Proteomic Analysis
4.4. TMT Labeling of Tryptic Peptides from Prostate Cancer Cell Lines
4.5. Chromatography Fractionation
4.6. Mass Spectrometric Analysis
4.7. Protein Expression Data Analysis
4.8. qRT-PCR Analysis
- FUT8-Forward: GACAGAACTGGTTCAGCGGAGA
- FUT8-Reverse: GCAGTAGACCACATGATGGAGC
- Actin-Forward: CACCATTGGCAATGAGCGGTTC
- Actin-Reverse: AGGTCTTTGCGGATGTCCACGT
- EGFR-Forward: AACACCCTGGTCTGGAAGTACG
- EGFR-Reverse: TCGTTGGACAGCCTTCAAGACC
4.9. Xenograft Animal Models
4.10. Immuno-Histochemical (IHC) Staining
4.11. Fluorescence Microscopy
4.12. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Höti, N.; Lih, T.-S.; Pan, J.; Zhou, Y.; Yang, G.; Deng, A.; Chen, L.; Dong, M.; Yang, R.-B.; Tu, C.-F.; et al. A Comprehensive Analysis of FUT8 Overexpressing Prostate Cancer Cells Reveals the Role of EGFR in Castration Resistance. Cancers 2020, 12, 468. https://doi.org/10.3390/cancers12020468
Höti N, Lih T-S, Pan J, Zhou Y, Yang G, Deng A, Chen L, Dong M, Yang R-B, Tu C-F, et al. A Comprehensive Analysis of FUT8 Overexpressing Prostate Cancer Cells Reveals the Role of EGFR in Castration Resistance. Cancers. 2020; 12(2):468. https://doi.org/10.3390/cancers12020468
Chicago/Turabian StyleHöti, Naseruddin, Tung-Shing Lih, Jianbo Pan, Yangying Zhou, Ganglong Yang, Ashely Deng, Lijun Chen, Mingmimg Dong, Ruey-Bing Yang, Cheng-Fen Tu, and et al. 2020. "A Comprehensive Analysis of FUT8 Overexpressing Prostate Cancer Cells Reveals the Role of EGFR in Castration Resistance" Cancers 12, no. 2: 468. https://doi.org/10.3390/cancers12020468