O-Linked N-Acetylglucosamine (O-GlcNAc) Expression Levels Epigenetically Regulate Colon Cancer Tumorigenesis by Affecting the Cancer Stem Cell Compartment via Modulating Expression of Transcriptional Factor MYBL1

被引:52
作者
Guo, Huabei [1 ]
Zhang, Bing [3 ,5 ]
Nairn, Alison V. [1 ]
Nagy, Tamas [2 ]
Moremen, Kelley W. [1 ]
Buckhaults, Phillip [4 ]
Pierce, Michael [1 ]
机构
[1] Univ Georgia, Dept Biochem & Mol Biol, Complex Carbohydrate Res Ctr, 315 Riverbend Rd, Athens, GA 30602 USA
[2] Univ Georgia, Coll Vet Med, Dept Pathol, Athens, GA 30602 USA
[3] Harvard Univ, Boston Childrens Hosp, Boston, MA 02115 USA
[4] Univ South Carolina, South Carolina Coll Pharm, Columbia, SC 29208 USA
[5] Jiaotong Univ, Shanghai Ctr Syst Biomed, 800 Dongchuan Rd, Shanghai 200240, Peoples R China
基金
美国国家卫生研究院;
关键词
EPITHELIAL-MESENCHYMAL TRANSITION; IN-VITRO PROPAGATION; A-MYB; DNA METHYLATION; GENE-EXPRESSION; TRANSFERASE OGT; HYPERMETHYLATION PROFILE; 3'-UNTRANSLATED REGION; UP-REGULATION; C-MYC;
D O I
10.1074/jbc.M116.763201
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
To study the regulation of colorectal adenocarcinoma progression by O-GlcNAc, we have focused on the O-GlcNAcmediated epigenetic regulation of human colon cancer stem cells (CCSC). Xenograft tumors from colon tumor cells with O-linked N-acetylglucosamine transferase (OGT) knockdown grew significantly slower than those formed from control cells, indicating a reduced proliferation of tumor cells due to inhibition ofOGTexpression. Significant reduction of the CCSC population was observed in the tumor cells after OGT knockdown, whereas tumor cells treated with the O-GlcNAcase inhibitor showed an increased CCSC population, indicating that O-GlcNAc levels regulated the CCSC compartment. When grown in suspension, tumor cells withOGTknockdown showed a reduced ability to form tumorspheres, indicating a reduced self-renewal of CCSC due to reduced levels of O-GlcNAc. ChIP-sequencing experiments using an anti-O-GlcNAc antibody revealed significant chromatin enrichment of O-GlcNAcmodified proteins at the promoter of the transcription factor MYBL1, which was also characterized by the presence of H3K27me3. RNA-sequencing analysis showed an increased expression of MYBL1 in tumor cells with OGT knockdown. Forced overexpression of MYBL1 led to a reduced population of CCSC and tumor growth in vivo, similar to the effects of OGT silencing. Moreover, two CpG islands near the transcription start site of MYBL1 were identified, and O-GlcNAc levels regulated their methylation status. These results strongly argue that O-GlcNAc epigenetically regulates MYBL1, functioning similarly to H3K27me3. The aberrant CCSC compartment observed after modulatingO-GlcNAc levels is therefore likely to result, at least in part, from the epigenetic regulation of MYBL1 expression by O-GlcNAc, thereby significantly affecting tumor progression.
引用
收藏
页码:4123 / 4137
页数:15
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