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

Overview

Journal ScientificWorldJournal

Publisher Wiley

Specialty Biology

Date 2014 Jan 25

PMID 24459426

Citations 8

Authors

Istvan Furi

Ferenc Sipos

Sandor Spisak

Gergo Kiszner

Barnabas Wichmann

Andrea Scholler

Zsolt Tulassay

Gyorgyi Muzes

Bela Molnar

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

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References

Klaschik S, Tross D, Klinman D . Inductive and suppressive networks regulate TLR9-dependent gene expression in vivo. J Leukoc Biol. 2009; 85(5):788-95. PMC: 2669411. DOI: 10.1189/jlb.1008671. View

Huidobro C, Urdinguio R, Rodriguez R, Mangas C, Calvanese V, Martinez-Camblor P . A DNA methylation signature associated with aberrant promoter DNA hypermethylation of DNMT3B in human colorectal cancer. Eur J Cancer. 2012; 48(14):2270-81. DOI: 10.1016/j.ejca.2011.12.019. View

Geiman T, Sankpal U, Robertson A, Chen Y, Mazumdar M, Heale J . Isolation and characterization of a novel DNA methyltransferase complex linking DNMT3B with components of the mitotic chromosome condensation machinery. Nucleic Acids Res. 2004; 32(9):2716-29. PMC: 419596. DOI: 10.1093/nar/gkh589. View

Ding W, Fang J, Chen X, Peng Y . The expression and clinical significance of DNA methyltransferase proteins in human gastric cancer. Dig Dis Sci. 2008; 53(8):2083-9. DOI: 10.1007/s10620-007-0145-2. View

Verthelyi D, Gursel M, Kenney R, Lifson J, Liu S, Mican J . CpG oligodeoxynucleotides protect normal and SIV-infected macaques from Leishmania infection. J Immunol. 2003; 170(9):4717-23. DOI: 10.4049/jimmunol.170.9.4717. View

Sipos F, Muzes G . Isolated lymphoid follicles in colon: switch points between inflammation and colorectal cancer?. World J Gastroenterol. 2011; 17(13):1666-73. PMC: 3072629. DOI: 10.3748/wjg.v17.i13.1666. View

Merrell M, Ilvesaro J, Lehtonen N, Sorsa T, Gehrs B, Rosenthal E . Toll-like receptor 9 agonists promote cellular invasion by increasing matrix metalloproteinase activity. Mol Cancer Res. 2006; 4(7):437-47. DOI: 10.1158/1541-7786.MCR-06-0007. View

Fang J, Cheng Z, Chen Y, Lu R, Yang L, Zhu H . Expression of Dnmt1, demethylase, MeCP2 and methylation of tumor-related genes in human gastric cancer. World J Gastroenterol. 2004; 10(23):3394-8. PMC: 4576216. DOI: 10.3748/wjg.v10.i23.3394. View

Fabbri M, Garzon R, Cimmino A, Liu Z, Zanesi N, Callegari E . MicroRNA-29 family reverts aberrant methylation in lung cancer by targeting DNA methyltransferases 3A and 3B. Proc Natl Acad Sci U S A. 2007; 104(40):15805-10. PMC: 2000384. DOI: 10.1073/pnas.0707628104. View

10.

Renbaum P, Abrahamove D, Fainsod A, Wilson G, Rottem S, Razin A . Cloning, characterization, and expression in Escherichia coli of the gene coding for the CpG DNA methylase from Spiroplasma sp. strain MQ1(M.SssI). Nucleic Acids Res. 1990; 18(5):1145-52. PMC: 330428. DOI: 10.1093/nar/18.5.1145. View

11.

Ewaschuk J, Backer J, Churchill T, Obermeier F, Krause D, Madsen K . Surface expression of Toll-like receptor 9 is upregulated on intestinal epithelial cells in response to pathogenic bacterial DNA. Infect Immun. 2007; 75(5):2572-9. PMC: 1865769. DOI: 10.1128/IAI.01662-06. View

12.

Lee J, Mo J, Katakura K, Alkalay I, Rucker A, Liu Y . Maintenance of colonic homeostasis by distinctive apical TLR9 signalling in intestinal epithelial cells. Nat Cell Biol. 2006; 8(12):1327-36. DOI: 10.1038/ncb1500. View

13.

Liang X, Moseman E, Farrar M, Bachanova V, Weisdorf D, Blazar B . Toll-like receptor 9 signaling by CpG-B oligodeoxynucleotides induces an apoptotic pathway in human chronic lymphocytic leukemia B cells. Blood. 2010; 115(24):5041-52. PMC: 2890142. DOI: 10.1182/blood-2009-03-213363. View

14.

Ilvesaro J, Merrell M, Swain T, Davidson J, Zayzafoon M, Harris K . Toll like receptor-9 agonists stimulate prostate cancer invasion in vitro. Prostate. 2007; 67(7):774-81. DOI: 10.1002/pros.20562. View

15.

Fischer M, Ehlers M . Toll-like receptors in autoimmunity. Ann N Y Acad Sci. 2008; 1143:21-34. DOI: 10.1196/annals.1443.012. View

16.

Schaefer T, DeSouza K, Fahey J, Beagley K, Wira C . Toll-like receptor (TLR) expression and TLR-mediated cytokine/chemokine production by human uterine epithelial cells. Immunology. 2004; 112(3):428-36. PMC: 1782499. DOI: 10.1111/j.1365-2567.2004.01898.x. View

17.

Akira S, Hemmi H . Recognition of pathogen-associated molecular patterns by TLR family. Immunol Lett. 2003; 85(2):85-95. DOI: 10.1016/s0165-2478(02)00228-6. View

18.

Klaschik S, Tross D, Shirota H, Klinman D . Short- and long-term changes in gene expression mediated by the activation of TLR9. Mol Immunol. 2009; 47(6):1317-24. PMC: 2830361. DOI: 10.1016/j.molimm.2009.11.014. View

19.

Gosu V, Basith S, Kwon O, Choi S . Therapeutic applications of nucleic acids and their analogues in Toll-like receptor signaling. Molecules. 2012; 17(11):13503-29. PMC: 6269001. DOI: 10.3390/molecules171113503. View

20.

Eads C, Danenberg K, Kawakami K, Saltz L, Danenberg P, Laird P . CpG island hypermethylation in human colorectal tumors is not associated with DNA methyltransferase overexpression. Cancer Res. 1999; 59(10):2302-6. View