Amplified and overexpressed gene in colorectal cancers
View Patent ↗This invention relates to methods to diagnose colon cancer and other proliferative diseases. Gene 26#77 is identified herein as a novel oncogene. Methods are provided for diagnosing and treating a disease or disorder characterized by amplification of the 26#77 gene and/or overexpression of 26#77 gene products. The 26#77 gene is located on chromosome 20q13.2, a region whose amplification is associated with a poor cancer prognosis. The 26#77 gene is amplified and 26#77 RNA and protein are overexpressed in 60% of colorectal cancers.
1. A method for determining the presence or absence of a colorectal cancer cell in a patient, the method comprising determining the level of a target nucleic acid that comprises a sequence at least 99% identical to SEQ ID NO: 1, wherein the level of the target nucleic acid is determined in a colorectal tissue sample from the patient and compared to the amount of the target nucleic acid in a reference sample, thereby determining the presence or absence of the colorectal cancer cell in the patient.
2. A method for determining the presence or absence of a colorectal cancer cell in a patient, the method comprising determining the level of a target nucleic acid that encodes the amino acid sequence of SEQ ID NO: 2, wherein the level of the target nucleic acid is determined in a colorectal tissue sample from the patient and compared to the amount of the target nucleic acid in a reference sample, thereby determining the presence or absence of the colorectal cancer cell in the patient.
3. The method of claim 1 or claim 2 , wherein the colorectal tissue sample comprises isolated nucleic acids.
4. The method of claim 3 , further comprising the step of amplifying nucleic acids before the step of determining the level of the nucleic acid.
5. The method of claim 3 , wherein the isolated nucleic acids are mRNA.
6. The method of claim 1 or claim 2 , wherein the step of determining the level of target nucleic acid is carried out using in situ hybridization.
7. The method of claim 1 or claim 2 , wherein the step of determining the level of target nucleic acid is carried out using a labeled nucleic acid probe that selectively hybridizes to SEQ ID NO: 1 under stringent hybridization conditions, wherein the stringent hybridization is carried out at 65° C. in a buffer comprising 5×SSC and 1% SDS, followed by a wash at 65° C. in a buffer comprising 0.2×SSC and 0.1% SDS; or is carried out at 42° C. in a buffer comprising 50% formamide, 5×SSC, and 1%SDS; followed by a wash at 65° C. in a buffer comprising 0.2×SSC and 0.1% SDS.
8. The method of claim 1 or claim 2 , wherein the step of determining the level of target nucleic acid is carried out using a nucleic acid probe immobilized to a solid support, wherein the probe selectively hybridizes to SEQ ID NO: 1 under stringent hybridization conditions, wherein the stringent hybridization is carried out at 65° C. in a buffer comprising 5×SSC and 1% SDS, followed by a wash at 65° C. in a buffer comprising 0.2×SSC and 0.1% SDS and 1% SDS; or is carried out at 42° C. in a buffer comprising 50% formamide, 5×SSC, and 1%SDS; followed by a wash at 65° C. in a buffer comprising 0.2×SSC and 0.1% SDS.
9. The method of claim 1 or claim 2 , wherein the step of determining the level of target nucleic acid is carried out using Northern blot analysis.
10. The method of claim 1 or claim 2 , wherein the reference sample is from normal colorectal tissue.
11. The method of claim 1 or claim 2 , wherein the patient is undergoing a therapeutic regimen to treat colorectal cancer.
12. The method of claim 1 or claim 2 , wherein the patient is suspected of having colorectal cancer.
13. A method for determining the presence or absence of a colorectal cancer cell in a patient, the method comprising determining the level of a target nucleic acid that comprises the sequence of SEQ ID NO: 1, wherein the level of the target nucleic acid is determined in a colorectal tissue sample from the patient and compared to the amount of the target nucleic acid in a reference sample, thereby determining the presence or absence of the colorectal cancer cell in the patient.
14. The method of claim 3 , wherein the isolated nucleic acids are DNA.
15. The method of claim 1 or claim 2 , wherein the step of determining the level of target nucleic acid is carried out using a micro array platform.