Abstract
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive tumor and still remains a challenge for its lack of effective therapeutic strategies, which is due to the late diagnosis of this disease. Methylation markers might improve early detection and surveillance of PDAC. Furthermore, analysis of hypermethylation in the tumor tissue might help to identify new targets for therapeutic intervention and improve the understanding of the pathophysiological changes occurring in pancreatic cancer. Methylation specific PCR is the method of choice if a small number of genes will be tested in a larger set of patient samples. After DNA isolation by standard procedure, the DNA is then modified using sodium bisulfite. This DNA can then be used in qualitative and quantitative PCR assays.
KB and MH contributed equallyl.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Jemal, A., Siegel, R., Xu, J., and Ward, E. (2010) Cancer statistics, 2010. CA Cancer J Clin 60, 277–300.
Schneider, G., Siveke, J.T., Eckel, F., and Schmid, R.M. (2005) Pancreatic cancer: basic and clinical aspects. Gastroenterology 128, 1606–1625.
Kulis, M. and Esteller, M. (2010) DNA methylation and cancer. Adv Genet 70, 27–56.
Nakhasi, H.L., Lynch, K.R., Dolan, K.P., Unterman, R.D., and Feigelson P. (1981) Covalent modification and repressed transcription of a gene in hepatoma cells. Proc Natl Acad Sci USA 78, 834–837.
Lofton-Day, C., Model, F., Devos, T., Tetzner, R., Distler, J., Schuster, M. et al (2008) DNA methylation biomarkers for blood-based colorectal cancer screening. Clin Chem 54, 414–423.
Tan, A.C., Jimeno, A., Lin, S.H., Wheelhouse, J., Chan, F., Solomon, A. et al (2009) Characterizing DNA methylation patterns in pancreatic cancer genome. Mol Oncol 3, 425–438.
Wehrum, D., Grutzmann, R., Hennig, M., Saeger, H.D., and Pilarsky, C. (2008) Recent patents concerning diagnostic and therapeutic applications of aberrantly methylated sequences in pancreatic cancer. Recent Pat DNA Gene Seq 2, 97–106.
Kuroki, T., Tajima, Y., and Kanematsu, T. (2004) Role of hypermethylation on carcinogenesis in the pancreas. Surg Today 34, 981–986.
Wright, D.K. and Manos, M.M. (1990). Sample preparation from paraffin-embedded tissues. In: Innis M.A., Gelfont D.H., Sninsky J.J. and White T.J. (eds). PCR Protocols: A guide to methods and applications. Academic Press, San Diego. pp 153–158.
Hong, S.M., Kelly, D., Griffith, M., Omura, N., Li, A., Li, C.P. et al. (2008). Multiple genes are hypermethylated in intraductal papillary mucinous neoplasms of the pancreas. Mod Pathol 12, 1499–1507.
Noetzel, E., Veeck, J., Niederacher, D., Galm, O., Horn, F., Hartmann, A. et al. (2008) Promoter methylation-associated loss of ID4 expression is a marker of tumour recurrence in human breast cancer. BMC Cancer 8, 154.
Okami, J., Simeone, D.M., and Logsdon, C.D. (2004) Silencing of the hypoxia-inducible cell death protein BNIP3 in pancreatic cancer. Cancer Res 64, 5338–5346.
Dunn, J.R., Panutsopulos, D., Shaw, M.W. Heighway, J., Dormer, R., Salmo, E.N. et al. (2004) METH-2 silencing and promoter hypermethylation in NSCLC. Br J Cancer 91, 1149–1154.
Acknowledgements
I am grateful to Beatrix Jahnke for performing DNA isolation and bisulfite modification in our lab. Thanks also to Alfred E. Neumann for fruitful discussions.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Biewusch, K., Heyne, M., Grützmann, R., Pilarsky, C. (2012). DNA Methylation in Pancreatic Cancer: Protocols for the Isolation of DNA and Bisulfite Modification. In: Dumitrescu, R., Verma, M. (eds) Cancer Epigenetics. Methods in Molecular Biology, vol 863. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-612-8_16
Download citation
DOI: https://doi.org/10.1007/978-1-61779-612-8_16
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-611-1
Online ISBN: 978-1-61779-612-8
eBook Packages: Springer Protocols