Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer
In colorectal cancer (CRC), upregulation of the C-X-C motif chemokine receptor 4 (CXCR4) is correlated with metastasis and poor prognosis, highlighting the need to further elucidate <i>CXCR4</i>’s regulation in CRC. For the first time, DNA methylation and 5-hydroxymethylcytosin...
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MDPI AG
2020-02-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/12/3/539 |
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| author | Alexei J. Stuckel Wei Zhang Xu Zhang Shuai Zeng Urszula Dougherty Reba Mustafi Qiong Zhang Elsa Perreand Tripti Khare Trupti Joshi Diana C. West-Szymanski Marc Bissonnette Sharad Khare |
| author_facet | Alexei J. Stuckel Wei Zhang Xu Zhang Shuai Zeng Urszula Dougherty Reba Mustafi Qiong Zhang Elsa Perreand Tripti Khare Trupti Joshi Diana C. West-Szymanski Marc Bissonnette Sharad Khare |
| author_sort | Alexei J. Stuckel |
| collection | DOAJ |
| description | In colorectal cancer (CRC), upregulation of the C-X-C motif chemokine receptor 4 (CXCR4) is correlated with metastasis and poor prognosis, highlighting the need to further elucidate <i>CXCR4</i>’s regulation in CRC. For the first time, DNA methylation and 5-hydroxymethylcytosine aberrations were investigated to better understand the epigenetic regulation of <i>CXCR4</i> in CRC. CXCR4 expression levels were measured using qPCR and immunoblotting in normal colon tissues, primary colon cancer tissues and CRC cell lines. Publicly available RNA-seq and methylation data from The Cancer Genome Atlas (TCGA) were extracted from tumors from CRC patients. The DNA methylation status spanning <i>CXCR4</i> gene was evaluated using combined bisulfite restriction analysis (COBRA). The methylation status in the <i>CXCR4</i> gene body was analyzed using previously performed nano-hmC-seal data from colon cancers and adjacent normal colonic mucosa. CXCR4 expression levels were significantly increased in tumor stromal cells and in tumor colonocytes, compared to matched cell types from adjacent normal-appearing mucosa. <i>CXCR4</i> promoter methylation was detected in a minority of colorectal tumors in the TCGA. The CpG island of the <i>CXCR4</i> promoter showed increased methylation in three of four CRC cell lines. CXCR4 protein expression differences were also notable between microsatellite stable (MSS) and microsatellite instable (MSI) tumor cell lines. While differential methylation was not detected in <i>CXCR4</i>, enrichment of 5-hydroxymethylcytosine (5hmC) in <i>CXCR4</i> gene bodies in CRC was observed compared to adjacent mucosa. |
| first_indexed | 2024-03-12T08:35:08Z |
| format | Article |
| id | doaj.art-b26079f500e64761aac767d1bd7fdef3 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T08:35:08Z |
| publishDate | 2020-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-b26079f500e64761aac767d1bd7fdef32023-09-02T17:20:19ZengMDPI AGCancers2072-66942020-02-0112353910.3390/cancers12030539cancers12030539Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal CancerAlexei J. Stuckel0Wei Zhang1Xu Zhang2Shuai Zeng3Urszula Dougherty4Reba Mustafi5Qiong Zhang6Elsa Perreand7Tripti Khare8Trupti Joshi9Diana C. West-Szymanski10Marc Bissonnette11Sharad Khare12Department of Medicine, Division of Gastroenterology and Hepatology, University of Missouri, Columbia, MO 65212, USADepartment of Preventive Medicine and The Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USADepartment of Medicine, University of Illinois, Chicago, IL, 60607, USABond Life Sciences Center, University of Missouri, Columbia, MO 65201, USADepartment of Medicine, Section of Gastroenterology, Hepatology and Nutrition, The University of Chicago, Chicago, IL 60637, USADepartment of Medicine, Section of Gastroenterology, Hepatology and Nutrition, The University of Chicago, Chicago, IL 60637, USADepartment of Medicine, Division of Gastroenterology and Hepatology, University of Missouri, Columbia, MO 65212, USADepartment of Medicine, Division of Gastroenterology and Hepatology, University of Missouri, Columbia, MO 65212, USADepartment of Medicine, Division of Gastroenterology and Hepatology, University of Missouri, Columbia, MO 65212, USABond Life Sciences Center, University of Missouri, Columbia, MO 65201, USADepartment of Medicine, Section of Gastroenterology, Hepatology and Nutrition, The University of Chicago, Chicago, IL 60637, USADepartment of Medicine, Section of Gastroenterology, Hepatology and Nutrition, The University of Chicago, Chicago, IL 60637, USADepartment of Medicine, Division of Gastroenterology and Hepatology, University of Missouri, Columbia, MO 65212, USAIn colorectal cancer (CRC), upregulation of the C-X-C motif chemokine receptor 4 (CXCR4) is correlated with metastasis and poor prognosis, highlighting the need to further elucidate <i>CXCR4</i>’s regulation in CRC. For the first time, DNA methylation and 5-hydroxymethylcytosine aberrations were investigated to better understand the epigenetic regulation of <i>CXCR4</i> in CRC. CXCR4 expression levels were measured using qPCR and immunoblotting in normal colon tissues, primary colon cancer tissues and CRC cell lines. Publicly available RNA-seq and methylation data from The Cancer Genome Atlas (TCGA) were extracted from tumors from CRC patients. The DNA methylation status spanning <i>CXCR4</i> gene was evaluated using combined bisulfite restriction analysis (COBRA). The methylation status in the <i>CXCR4</i> gene body was analyzed using previously performed nano-hmC-seal data from colon cancers and adjacent normal colonic mucosa. CXCR4 expression levels were significantly increased in tumor stromal cells and in tumor colonocytes, compared to matched cell types from adjacent normal-appearing mucosa. <i>CXCR4</i> promoter methylation was detected in a minority of colorectal tumors in the TCGA. The CpG island of the <i>CXCR4</i> promoter showed increased methylation in three of four CRC cell lines. CXCR4 protein expression differences were also notable between microsatellite stable (MSS) and microsatellite instable (MSI) tumor cell lines. While differential methylation was not detected in <i>CXCR4</i>, enrichment of 5-hydroxymethylcytosine (5hmC) in <i>CXCR4</i> gene bodies in CRC was observed compared to adjacent mucosa.https://www.mdpi.com/2072-6694/12/3/539colorectal cancerdna methylationepigenetic regulationcxcr4 gene expression5-hydroxymethylcytosine |
| spellingShingle | Alexei J. Stuckel Wei Zhang Xu Zhang Shuai Zeng Urszula Dougherty Reba Mustafi Qiong Zhang Elsa Perreand Tripti Khare Trupti Joshi Diana C. West-Szymanski Marc Bissonnette Sharad Khare Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer Cancers colorectal cancer dna methylation epigenetic regulation cxcr4 gene expression 5-hydroxymethylcytosine |
| title | Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer |
| title_full | Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer |
| title_fullStr | Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer |
| title_full_unstemmed | Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer |
| title_short | Enhanced CXCR4 Expression Associates with Increased Gene Body 5-Hydroxymethylcytosine Modification but not Decreased Promoter Methylation in Colorectal Cancer |
| title_sort | enhanced cxcr4 expression associates with increased gene body 5 hydroxymethylcytosine modification but not decreased promoter methylation in colorectal cancer |
| topic | colorectal cancer dna methylation epigenetic regulation cxcr4 gene expression 5-hydroxymethylcytosine |
| url | https://www.mdpi.com/2072-6694/12/3/539 |
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