Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation Status
To understand the biologic role of self-DNA bound to Toll-like Receptor 9 (TLR9), we assayed its effect on gene and methyltransferase expressions and cell differentiation in HT29 cells. HT29 cells were incubated separately with type-1 (normally methylated/nonfragmented), type-2 (normally methylated/...
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| Format: | Article |
| Language: | English |
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Wiley
2013-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2013/293296 |
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| author | István Fűri Ferenc Sipos Sándor Spisák Gergő Kiszner Barnabás Wichmann Andrea Schöller Zsolt Tulassay Györgyi Műzes Béla Molnár |
| author_facet | István Fűri Ferenc Sipos Sándor Spisák Gergő Kiszner Barnabás Wichmann Andrea Schöller Zsolt Tulassay Györgyi Műzes Béla Molnár |
| author_sort | István Fűri |
| collection | DOAJ |
| description | To understand the biologic role of self-DNA bound to Toll-like Receptor 9 (TLR9), we assayed its effect on gene and methyltransferase expressions and cell differentiation in HT29 cells. HT29 cells were incubated separately with type-1 (normally methylated/nonfragmented), type-2 (normally methylated/fragmented), type-3 (hypermethylated/nonfragmented), or type-4 (hypermethylated/fragmented) self-DNAs. Expression levels of TLR9-signaling and proinflammatory cytokine-related genes were assayed by qRT-PCR. Methyltransferase activity and cell differentiation were examined by using DNA methyltransferase (DNMT1, -3A, -3B) and cytokeratin (CK) antibodies. Treatment with type-1 DNA resulted in significant increase in TLR9 expression. Type-2 treatment resulted in the overexpression of TLR9-related signaling molecules (MYD88A, TRAF6) and the IL8 gene. In the case of type-3 treatment, significant overexpression of NFkB, IRAK2, and IL8 as well as downregulation of TRAF6 was detected. Using type-4 DNA, TRAF6 and MYD88A gene expression was upregulated, while MYD88B, IRAK2, IL8, and TNFSF10 were all underexpressed. CK expression was significantly higher only after type-1 DNA treatment. DNMT3A expression could also be induced by type-1 DNA treatment. DNA structure may play a significant role in activation of the TLR9-dependent and even independent proinflammatory pathways. There may be a molecular link between TLR9 signaling and DNMT3A. The mode of self-DNA treatment may influence HT29 cell differentiation. |
| format | Article |
| id | doaj-art-2adea9b98aff4bed812781a7a62d4f05 |
| institution | Kabale University |
| issn | 1537-744X |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-2adea9b98aff4bed812781a7a62d4f052025-02-03T01:31:25ZengWileyThe Scientific World Journal1537-744X2013-01-01201310.1155/2013/293296293296Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation StatusIstván Fűri0Ferenc Sipos1Sándor Spisák2Gergő Kiszner3Barnabás Wichmann4Andrea Schöller5Zsolt Tulassay6Györgyi Műzes7Béla Molnár82nd Department of Internal Medicine, Semmelweis University, Szentkirályi Street 46, Budapest 1088, Hungary2nd Department of Internal Medicine, Semmelweis University, Szentkirályi Street 46, Budapest 1088, HungaryMolecular Medicine Research Unit, Hungarian Academy of Sciences, Nádor Street 7, Budapest 1051, Hungary1st Department of Pathology and Experimental Oncology, Semmelweis University, Üllői Street 26, Budapest 1085, Hungary2nd Department of Internal Medicine, Semmelweis University, Szentkirályi Street 46, Budapest 1088, Hungary2nd Department of Internal Medicine, Semmelweis University, Szentkirályi Street 46, Budapest 1088, HungaryMolecular Medicine Research Unit, Hungarian Academy of Sciences, Nádor Street 7, Budapest 1051, Hungary2nd Department of Internal Medicine, Semmelweis University, Szentkirályi Street 46, Budapest 1088, HungaryMolecular Medicine Research Unit, Hungarian Academy of Sciences, Nádor Street 7, Budapest 1051, HungaryTo understand the biologic role of self-DNA bound to Toll-like Receptor 9 (TLR9), we assayed its effect on gene and methyltransferase expressions and cell differentiation in HT29 cells. HT29 cells were incubated separately with type-1 (normally methylated/nonfragmented), type-2 (normally methylated/fragmented), type-3 (hypermethylated/nonfragmented), or type-4 (hypermethylated/fragmented) self-DNAs. Expression levels of TLR9-signaling and proinflammatory cytokine-related genes were assayed by qRT-PCR. Methyltransferase activity and cell differentiation were examined by using DNA methyltransferase (DNMT1, -3A, -3B) and cytokeratin (CK) antibodies. Treatment with type-1 DNA resulted in significant increase in TLR9 expression. Type-2 treatment resulted in the overexpression of TLR9-related signaling molecules (MYD88A, TRAF6) and the IL8 gene. In the case of type-3 treatment, significant overexpression of NFkB, IRAK2, and IL8 as well as downregulation of TRAF6 was detected. Using type-4 DNA, TRAF6 and MYD88A gene expression was upregulated, while MYD88B, IRAK2, IL8, and TNFSF10 were all underexpressed. CK expression was significantly higher only after type-1 DNA treatment. DNMT3A expression could also be induced by type-1 DNA treatment. DNA structure may play a significant role in activation of the TLR9-dependent and even independent proinflammatory pathways. There may be a molecular link between TLR9 signaling and DNMT3A. The mode of self-DNA treatment may influence HT29 cell differentiation.http://dx.doi.org/10.1155/2013/293296 |
| spellingShingle | István Fűri Ferenc Sipos Sándor Spisák Gergő Kiszner Barnabás Wichmann Andrea Schöller Zsolt Tulassay Györgyi Műzes Béla Molnár Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation Status The Scientific World Journal |
| title | Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation Status |
| title_full | Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation Status |
| title_fullStr | Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation Status |
| title_full_unstemmed | Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation Status |
| title_short | Association of Self-DNA Mediated TLR9-Related Gene, DNA Methyltransferase, and Cytokeratin Protein Expression Alterations in HT29-Cells to DNA Fragment Length and Methylation Status |
| title_sort | association of self dna mediated tlr9 related gene dna methyltransferase and cytokeratin protein expression alterations in ht29 cells to dna fragment length and methylation status |
| url | http://dx.doi.org/10.1155/2013/293296 |
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