Methods and nucleic acids for determining the prognosis of a cancer subject
The invention provides methods, nucleic acids and kits for determining the prognosis of a subject having cancer. The invention discloses genomic sequences the methylation patterns of which have utility for the improved detection of said disorder, thereby enabling the improved diagnosis and treatment of patients.
1. A method for determining methylation of SEPTIN-9 (SEQ ID NO: 1) genomic colon or colorectal tumor cell DNA of a human subject with colorectal cancer following removal of a primary colon or colorectal tumor but prior to further treatment, the method comprising:
detecting the presence of, or measuring the level of, methylated genomic SEPTIN-9 DNA in a stool, blood, serum or plasma sample obtained from the human subject following the removal of the primary colon or colorectal tumor but prior to further treatment, wherein the methylated genomic SEPTIN-9 DNA is from colon or colorectal tumor cells, and wherein the detection uses a primer pair in which one of the primers hybridizes under stringent conditions to at least 9 nucleotides of the nucleic acid sequence of SEQ ID NO:26 or a complement thereof, and the other primer of the primer pair hybridizes under stringent conditions to at least 9 nucleotides of the nucleic acid sequence of SEQ ID NO:25 or a complement thereof, thereby
determining methylation of the SEPTIN-9 genomic colon or colorectal tumor cell DNA of the human subject after removal of the primary colon or colorectal tumor,
wherein the colorectal cancer is Stage III according to the Tumor-Node-Metastasis (TNM) staging system and the removal of the primary colon or colorectal tumor includes nearby lymph nodes.
2. The method of claim 1 , wherein one of the primers of the primer pair comprises the nucleic acid sequence of SEQ ID NO: 26, or a complement thereof, and the other primer of the primer pair hybridizes under stringent conditions to at least 9 nucleotides of the nucleic acid sequence of SEQ ID NO: 25 or a complement thereof.
3. The method of claim 1 , further comprising using a blocker that hybridizes under stringent conditions to at least 9 nucleotides of the nucleic acid sequence of SEQ ID NO: 27.
4. The method of claim 3 , further comprising using a probe that hybridizes under stringent conditions to at least 9 nucleotides of the nucleic acid sequence of SEQ ID NO: 28.
5. The method of claim 1 , wherein at least one primer of the primer pair comprises SEQ ID NO: 26, or a complement thereof.
6. The method of claim 1 , wherein at least one primer of the primer pair comprises SEQ ID NO: 25, or a complement thereof.
7. The method of claim 1 , wherein determining methylation of the SEPTIN-9 genomic colon or colorectal tumor cell DNA further comprises using primers consisting of SEQ ID NOs: 25-26, a blocker consisting of SEQ ID NO: 27, and a probe consisting of SEQ ID NO: 28.
8. The method of claim 4 , wherein the presence of methylated genomic SEPTIN-9 DNA in the stool, blood, serum or plasma sample following the removal of the primary colon or colorectal tumor, but prior to further treatment, indicates that the colorectal cancer of the human subject is aggressive.