Abstract
Interleukin-6 (IL-6) is overexpressed and contributes to tumor cell growth in cholangiocarcinoma. Enforced IL-6 production can alter the expression of specific microRNAs (miRNAs) involved in tumor growth, and moreover can modulate expression of methylation-dependent genes. Thus, we assessed the methylation-dependent regulation of miRNA expression in human malignant cholangiocytes stably transfected to overexpress IL-6. The expression of the methyltransferases DNA methyltransferase enzyme-1 and HASJ4442 was increased by IL-6 overexpression, but was decreased by the methylation inhibitor 5-aza-2′-deoxycytidine (5-aza-CdR). Expression profiling identified seven miRNAs that were significantly downregulated by IL-6 overexpression (<0.4-fold) and upregulated (>2-fold) by 5-aza-CdR. One of these, miR-370, is embedded in a CpG island. Although 5-aza-CdR increased miR-370 expression by 2.1-fold in malignant cells, the expression in nonmalignant cells was unchanged. The oncogene mitogen-activated protein kinase kinase kinase 8 (MAP3K8) was identified as a target of miR-370, and its expression was decreased by 5-aza-CdR in cholangiocarcinoma cells. Overexpression of IL-6 reduced miR-370 expression and reinstated MAP3K8 expression in vitro as well as in tumor cell xenografts in vivo. Thus, IL-6 may contribute to tumor growth by modulation of expression of selected miRNAs, such as miR-370. These studies define a mechanism by which inflammation-associated cytokines can epigenetically modulate gene expression and directly contribute to tumor biology.
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Abbreviations
- 5-aza-CdR:
-
5-aza-2′deoxycytidine
- DNMT:
-
DNA methyltransferase enzyme
- IBEC:
-
intrahepatic biliary epithelial cells
- IL-6:
-
interleukin-6
- MAP3K8:
-
mitogen-activated protein kinase kinase kinase 8
- miRNA:
-
microRNA
References
Aoki M, Hamada F, Sugimoto T, Sumida S, Akiyama T, Toyoshima K . (1993). The human cot proto-oncogene encodes two protein serine/threonine kinases with different transforming activities by alternative initiation of translation. J Biol Chem 268: 22723–22732.
Bandres E, Cubedo E, Agirre X, Malumbres R, Zarate R, Ramirez N et al. (2006). Identification by Real-time PCR of 13 mature microRNAs differentially expressed in colorectal cancer and non-tumoral tissues. Mol Cancer 5: 29.
Banerjee A, Gugasyan R, McMahon M, Gerondakis S . (2006). Diverse toll-like receptors utilize Tpl2 to activate extracellular signal-regulated kinase (ERK) in hemopoietic cells. Proc Natl Acad Sci USA 103: 3274–3279.
Baylin SB . (2005). DNA methylation and gene silencing in cancer. Nat Clin Pract Oncol 2 (Suppl 1): S4–S11.
Calin GA, Croce CM . (2006a). MicroRNA signatures in human cancers. Nat Rev Cancer 6: 857–866.
Calin GA, Croce CM . (2006b). MicroRNAs and chromosomal abnormalities in cancer cells. Oncogene 25: 6202–6210.
Chiariello M, Marinissen MJ, Gutkind JS . (2000). Multiple mitogen-activated protein kinase signaling pathways connect the cot oncoprotein to the c-jun promoter and to cellular transformation. Mol Cell Biol 20: 1747–1758.
Eliopoulos AG, Davies C, Blake SS, Murray P, Najafipour S, Tsichlis PN et al. (2002). The oncogenic protein kinase Tpl-2/Cot contributes to Epstein-Barr virus-encoded latent infection membrane protein 1-induced NF-kappaB signaling downstream of TRAF2. J Virol 76: 4567–4579.
Heinrich PC, Behrmann I, Haan S, Hermanns HM, Muller-Newen G, Schaper F . (2003). Principles of interleukin (IL)-6-type cytokine signalling and its regulation. Biochem J 374 (Part 1): 1–20.
Hodge DR, Peng B, Cherry JC, Hurt EM, Fox SD, Kelley JA et al. (2005). Interleukin 6 supports the maintenance of p53 tumor suppressor gene promoter methylation. Cancer Res 65: 4673–4682.
Hodge DR, Xiao W, Clausen PA, Heidecker G, Szyf M, Farrar WL . (2001). Interleukin-6 regulation of the human DNA methyltransferase (HDNMT) gene in human erythroleukemia cells. J Biol Chem 276: 39508–39511.
John B, Enright AJ, Aravin A, Tuschl T, Sander C, Marks DS . (2004). Human microRNA targets. PLoS Biol 2: E363.
Kiriakidou M, Nelson PT, Kouranov A, Fitziev P, Bouyioukos C, Mourelatos Z et al. (2004). A combined computational-experimental approach predicts human microRNA targets. Genes Dev 18: 1165–1178.
Krek A, Grun D, Poy MN, Wolf R, Rosenberg L, Epstein EJ et al. (2005). Combinatorial microRNA target predictions. Nat Genet 37: 495–500.
Lewis BP, Burge CB, Bartel DP . (2005). Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120: 15–20.
Makris A, Patriotis C, Bear SE, Tsichlis PN . (1993). Genomic organization and expression of Tpl-2 in normal cells and Moloney murine leukemia virus-induced rat T-cell lymphomas: activation by provirus insertion. J Virol 67: 4283–4289.
Meng F, Henson R, Lang M, Wehbe H, Maheshwari S, Mendell JT et al. (2006a). Involvement of human micro-RNA in growth and response to chemotherapy in human cholangiocarcinoma cell lines. Gastroenterology 130: 2113–2129.
Meng F, Yamagiwa Y, Taffetani S, Han J, Patel T . (2005). IL-6 activates serum and glucocorticoid kinase via p38alpha mitogen-activated protein kinase pathway. Am J Physiol Cell Physiol 289: C971–C981.
Meng F, Yamagiwa Y, Ueno Y, Patel T . (2006b). Over-expression of interleukin-6 enhances cell survival and transformed cell growth in human malignant cholangiocytes. J Hepatol 44: 1055–1065.
Ohara R, Hirota S, Onoue H, Nomura S, Kitamura Y, Toyoshima K . (1995). Identification of the cells expressing cot proto-oncogene mRNA. J Cell Sci 108 (Part 1): 97–103.
Okada K, Shimizu Y, Nambu S, Higuchi K, Watanabe A . (1994). Interleukin-6 functions as an autocrine growth factor in a cholangiocarcinoma cell line. J Gastroenterol Hepatol 9: 462–467.
Park J, Tadlock L, Gores GJ, Patel T . (1999). Inhibition of interleukin 6-mediated mitogen-activated protein kinase activation attenuates growth of a cholangiocarcinoma cell line. Hepatology 30: 1128–1133.
Patel T . (2006). Cholangiocarcinoma. Nat Clin Pract Gastroenterol Hepatol 3: 33–42.
Patriotis C, Makris A, Bear SE, Tsichlis PN . (1993). Tumor progression locus 2 (Tpl-2) encodes a protein kinase involved in the progression of rodent T-cell lymphomas and in T-cell activation. Proc Natl Acad Sci USA 90: 2251–2255.
Saeed AI, Sharov V, White J, Li J, Liang W, Bhagabati N et al. (2003). TM4: a free, open-source system for microarray data management and analysis. Biotechniques 34: 374–378.
Saito Y, Liang G, Egger G, Friedman JM, Chuang JC, Coetzee GA et al. (2006). Specific activation of microRNA-127 with downregulation of the proto-oncogene BCL6 by chromatin-modifying drugs in human cancer cells. Cancer Cell 9: 435–443.
Sourvinos G, Tsatsanis C, Spandidos DA . (1999). Overexpression of the Tpl-2/Cot oncogene in human breast cancer. Oncogene 18: 4968–4973.
Tadlock L, Patel T . (2001). Involvement of p38 mitogen-activated protein kinase signaling in transformed growth of a cholangiocarcinoma cell line. Hepatology 33: 43–51.
Takai D, Jones PA . (2002). Comprehensive analysis of CpG islands in human chromosomes 21 and 22. Proc Natl Acad Sci USA 99: 3740–3745.
Wehbe H, Henson R, Meng F, Mize-Berge J, Patel T . (2006). Interleukin-6 contributes to growth in cholangiocarcinoma cells by aberrant promoter methylation and gene expression. Cancer Res 66: 10517–10524.
Yamagiwa Y, Marienfeld C, Tadlock L, Patel T . (2003). Translational regulation by p38 mitogen-activated protein kinase signaling during human cholangiocarcinoma growth. Hepatology 38: 158–166.
Acknowledgements
This study was supported by grants DK069370 from the National Institutes of Health.
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Meng, F., Wehbe-Janek, H., Henson, R. et al. Epigenetic regulation of microRNA-370 by interleukin-6 in malignant human cholangiocytes. Oncogene 27, 378–386 (2008). https://doi.org/10.1038/sj.onc.1210648
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DOI: https://doi.org/10.1038/sj.onc.1210648
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