Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer
Abstract
:1. Introduction
2. Results
2.1. CXCR4 Is Highly-Expressed in Colorectal Cancers
2.2. CXCR4 Promoter Hypomethylation Is Associated with Metastatic CRC
2.3. CXCR4 Promoter Hypermethylation Correlates with Decreased Expression in CRC Cell Lines
2.4. Increased 5hmC Deposition within CXCR4 Gene Body Correlates with Increased CXCR4 Expression
3. Discussion
4. Materials and Methods
4.1. Human Tissue Collection
4.2. Isolation of Coloncytes and Stromal Cells from Colonic Mucosa
4.3. Cell Culture
4.4. Measurement of CXCR4 Steady-State Transcript Levels
4.5. CXCR4 Western Blotting
4.6. Combined Bisulfite Restriction Analysis
4.7. 5hmC-Modified Locations in the CXCR4 Gene Body
4.8. Inhibition of 5mC Deposition in Colorectal Cancer Cell Lines HCT116 and HT29
4.9. CXCR4 Bioinformatics Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Stuckel, A.J.; Zhang, W.; Zhang, X.; Zeng, S.; Dougherty, U.; Mustafi, R.; Zhang, Q.; Perreand, E.; Khare, T.; Joshi, T.; et al. Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer. Cancers 2020, 12, 539. https://doi.org/10.3390/cancers12030539
Stuckel AJ, Zhang W, Zhang X, Zeng S, Dougherty U, Mustafi R, Zhang Q, Perreand E, Khare T, Joshi T, et al. Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer. Cancers. 2020; 12(3):539. https://doi.org/10.3390/cancers12030539
Chicago/Turabian StyleStuckel, Alexei J., Wei Zhang, Xu Zhang, Shuai Zeng, Urszula Dougherty, Reba Mustafi, Qiong Zhang, Elsa Perreand, Tripti Khare, Trupti Joshi, and et al. 2020. "Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer" Cancers 12, no. 3: 539. https://doi.org/10.3390/cancers12030539