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DNA methylation, inflammation and cancer
A selection of talks on Cell Biology
Gene structure, expression and regulation: DNA structure and replication
- Dr. Carole Sargent
- University of Cambridge, UK
Preclinical translation of mesenchymal stem cell therapies
- Dr. Peter Childs
- University of Strathclyde, UK
My name is Gerd Pfeiffer. The topic of my talk is DNA methylation, inflammation, and cancer.
The talk will be divided into four parts, in part one, a method for DNA methylation analysis, genome wide will be described. In the second part, data on the DNA methylation patterns in B cells will be reported. In part three, I will discuss our data on the inner methylation in lung cancer. And in the last part, the relationship between inflammation, DNA methylation and cancer and targeting of specific genes will be discussed.
DNA methylation patterns in mammalian cells are generally quite stable. During DNA replication, methylation at CpG sites is copied by DNA methyltransferase one. Methylation can also be introduced de novo or methylation can be lost by DNA demethylation processes, either active or passive DNA demethylation. These mechanisms can be altered in diseased tissue.
There's many different methods to analyze DNA methylation on a genome-wide scale. Some are based on methylation sensitive restriction enzymes. They're shown here in panels A, B and C, the information can be detected by two dimensional gel electrophoresis by PCR or by micro arrays. Another method used is an antibody against methylated cytosine is shown here in panel D and it's called Methyl-DIP. This method is like immunoprecipitation and the methylated immunoprecipitated DNA can be analyzed either by high throughput sequencing or by microarray chip analysis. Another way to look at methylated genes is to use a methylation inhibitor, 5-Aza-cytidine or DAC. Genes that become reactivated by DAC treatment shown here in panel E and then be scored and methylation must then be confirmed by different, more specific methods. We developed a method for genome wide DNA methylation profiling,