Abstract
Pancreatic ductal adenocarcinoma (PDAC) is characterized by an abundant stroma enriched with hyaluronan (HA), a major component of extracellular matrix known to play a critical role in tumor progression. The mechanisms that regulate HA synthesis in PDAC are poorly understood. To investigate whether DNA methylation and HA production from PDAC cells are associated, we studied the effect of 5-aza-2′-deoxycitidine (5-aza-dC), an inhibitor of DNA methylation, or DNA methyltransferase 1 (DNMT1) knockdown by small interfering RNA, on the HA production from PDAC cells. HA production into the conditioned medium was evaluated in PDAC cells treated with 5-aza-dC or DNMT1 knockdown. mRNA expression of HA synthase (HAS) genes was investigated by real-time RT-PCR. Treatment of PDAC cells with 5-aza-dC led to a significant increase in the HA production (up to 2.5-fold increase) in all 4 cell lines tested. This enhanced HA production by 5-aza-dC treatment was accompanied by increased mRNA expression of HAS2 and HAS3. Furthermore, increased HA production and HAS2/HAS3 mRNA expression was also observed in PDAC cells by knockdown of DNMT1. These findings provide evidence, for the first time, that epigenetic mechanism is involved in the regulation of HA synthesis in PDAC cells.
Abbreviations
- PDAC:
-
Pancreatic ductal adenocarcinoma
- ECM:
-
Extracellular matrix
- HA:
-
Hyaluronan
- MMP:
-
Matrix metalloproteinase
- 5-aza-dC:
-
5-aza-2′-deoxycytidine
- DNMT1:
-
DNA methyltransferase 1
- HAS:
-
HA synthase
- HPDE:
-
Human pancreatic duct epithelial cell line
References
Bardeesy N, DePinho RA (2002) Pancreatic cancer biology and genetics. Nat Rev Cancer 2(12):897–909
Hidalgo M (2010) Pancreatic cancer. N Engl J Med 362(17):1605–1617
Vincent A et al (2011) Pancreatic cancer. Lancet 378(9791):607–620
Sato N, Goggins M (2006) The role of epigenetic alterations in pancreatic cancer. J Hepatobiliary Pancreat Surg 13(4):286–295
Mahadevan D, Von Hoff DD (2007) Tumor-stroma interactions in pancreatic ductal adenocarcinoma. Mol Cancer Ther 6(4):1186–1197
Erkan M et al (2010) Tumor microenvironment and progression of pancreatic cancer. Exp Oncol 32(3):128–131
Toole BP (2004) Hyaluronan: from extracellular glue to pericellular cue. Nat Rev Cancer 4(7):528–539
Itano N, Zhuo L, Kimata K (2008) Impact of the hyaluronan-rich tumor microenvironment on cancer initiation and progression. Cancer Sci 99(9):1720–1725
Cheng XB et al (2013) Prognostic impact of hyaluronan and its regulators in pancreatic ductal adenocarcinoma. PLoS ONE 8(11):e80765
Provenzano PP et al (2012) Enzymatic targeting of the stroma ablates physical barriers to treatment of pancreatic ductal adenocarcinoma. Cancer Cell 21(3):418–429
Jacobetz MA et al (2012) Hyaluronan impairs vascular function and drug delivery in a mouse model of pancreatic cancer. Gut 62(1):112–120
Sato N et al (2003) Effects of 5-aza-2′-deoxycytidine on matrix metalloproteinase expression and pancreatic cancer cell invasiveness. J Natl Cancer Inst 95(4):327–330
Sato N et al (2003) SPARC/osteonectin is a frequent target for aberrant methylation in pancreatic adenocarcinoma and a mediator of tumor-stromal interactions. Oncogene 22(32):5021–5030
Sato N et al (2006) Differential and epigenetic gene expression profiling identifies frequent disruption of the RELN pathway in pancreatic cancers. Gastroenterology 130(2):548–565
Sato N et al (2003) Discovery of novel targets for aberrant methylation in pancreatic carcinoma using high-throughput microarrays. Cancer Res 63(13):3735–3742
Fries H et al (1994) Localisation of hyaluronate (HA) in primary tumors and nude mouse xenografts of human pancreatic carcinomas using a biotinylated HA-binding protein. Virchows Arch 424(1):7–12
Mahlbacher V et al (1992) Hyaluronan is a secretory product of human pancreatic adenocarcinoma cells. Eur J Cell Biol 58(1):28–34
Theocharis AD et al (2000) Pancreatic carcinoma is characterized by elevated content of hyaluronan and chondroitin sulfate with altered disaccharide composition. Biochim Biophys Acta 1502(2):201–206
Kultti A et al (2014) Accumulation of extracellular hyaluronan by hyaluronan synthase 3 promotes tumor growth and modulates the pancreatic cancer microenvironment. Biomed Res Int 2014:817613
Chamberlain AA et al (2014) DNA methylation is developmentally regulated for genes essential for cardiogenesis. J Am Heart Assoc 3(3):e000976
Jiang D, Liang J, Noble PW (2007) Hyaluronan in tissue injury and repair. Annu Rev Cell Dev Biol 23:435–461
Issa JP, Kantarjian HM (2009) Targeting DNA methylation. Clin Cancer Res 15(12):3938–3946
Yamada T et al (1996) Induction of Ley antigen by 5-aza-2′-deoxycytidine in association with differentiation and apoptosis in human pancreatic cancer cells. Anticancer Res 16(2):735–740
Shakya R et al (2013) Hypomethylating therapy in an aggressive stroma-rich model of pancreatic carcinoma. Cancer Res 73(2):885–896
Ateeq B et al (2008) Pharmacological inhibition of DNA methylation induces proinvasive and prometastatic genes in vitro and in vivo. Neoplasia 10(3):266–278
Poplineau M et al (2013) The DNA hypomethylating agent, 5-aza-2′-deoxycytidine, enhances tumor cell invasion through a transcription-dependent modulation of MMP-1 expression in human fibrosarcoma cells. Mol Carcinog 54(1):24–34
Ricciardelli C et al (2013) Chemotherapy-induced hyaluronan production: a novel chemoresistance mechanism in ovarian cancer. BMC Cancer 13:476
Acknowledgments
We thank Ms. Ueda (Research Assistant, Department of Surgery1, University of Occupational and Environmental Health, Kitakyushu, JAPAN) for her technical assistance.
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Kohi, S., Sato, N., Cheng, XB. et al. A novel epigenetic mechanism regulating hyaluronan production in pancreatic cancer cells. Clin Exp Metastasis 33, 225–230 (2016). https://doi.org/10.1007/s10585-015-9771-9
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DOI: https://doi.org/10.1007/s10585-015-9771-9