Methods for identifying cancer risk
Provided herein are tissue-specific differential methylated regions (T-DMRs) and cancer-related differential methylated regions (C-DMRs) and methods of use thereof. In one embodiment of the invention, there are provided methods of detecting a cell proliferative disorder by detecting altered methylation in one or more DMRs identified herein. In another embodiment of the invention, there are provided methods of determining clinical outcome by detecting altered methylation in one or more DMRs identified herein.
1. A method of detecting colon cancer in a subject comprising:
a) determining a methylation status of nucleic acid sequences in a sample from a tissue suspected of having colon cancer and a sample from a normal tissue of the subject,
wherein determining the methylation status comprises:
i) contacting the nucleic acid sequences of each sample with a plurality of control probes that selectively hybridize to genomic regions without CpG regions;
ii) generating M values for the plurality of control probes for each sample;
iii) calculating an averaged M value for each sample using the generated M values to define a value for unmethylated nucleic acid sequences for each sample;
iv) contacting the nucleic acid sequences of each sample with a second plurality of probes that selectively hybridize to genomic regions with CpG regions;
v) generating M values for the second plurality of probes and comparing the generated M values to the average M value for each sample to determine the methylation status of the nucleic acid sequences of each sample; and
vi) comparing the methylation status of each sample to identify cancer-specific differential methylated regions (C-DMRs) in the sample from the tissue suspected of having colon cancer, wherein the presence of C-DMRs in the sample from the tissue suspected of having colon cancer is indicative of colon cancer, thereby detecting colon cancer in the subject; and
b) administering a cancer therapy to the subject.
2. The method of claim 1 , wherein the nucleic acid sequences are within a gene.
3. The method of claim 1 , wherein the nucleic acid sequences are upstream or downstream of a gene.
4. The method of claim 1 , wherein the methylation status is hypomethylation.
5. The method of claim 1 , wherein the methylation status is hypermethylation.
6. The method of claim 1 , further comprising performing one or more techniques selected from the group consisting of a nucleic acid amplification, polymerase chain reaction (PCR), methylation specific PCR, bisulfite pyrosequencing, single-strand conformation polymorphism (SSCP) analysis, restriction analysis, microarray technology, and proteomics.