GRADING OF BREAST CANCER
Methods and compositions for the identification of breast cancer grade signatures are provided. The signature profiles are identified based upon multiple sampling of reference breast tissue samples from independent cases of breast cancer and provide a reliable set of molecular criteria for identification of cells as being in one or more particular stages and/or grades of breast cancer.
1 . A method to determine the grade of breast cancer cells in a sample from a human subject, said method comprising
assaying said sample for gene expression that is increased in a high proliferation index of grade III relative to grade I breast cancer, wherein said gene expression is assayed by synthesis of cDNA.
2 . The method of claim 1 , wherein the gene expression is assayed by synthesis of cDNA from expressed RNA of the gene CENPA (centromere protein A (17 kD), chromosomal location 2p24-p21).
3 . A method to determine the grade of breast cancer cells in a sample from a human subject, said method comprising
assaying said sample for gene expression that is increased in grade III relative to grade I breast cancer, wherein said gene expression is assayed by synthesis of cDNA from expressed RNA of a gene selected from
RACGAP1 (Rac GTPase activating protein 1, chromosomal location 12p13.2-p13.1),
RRM2 (ribonucleotide reductase M2 polypeptide, chromosomal location 2p25-p24), and
NEK2 (NIMA (never in mitosis gene a)-related kinase, chromosomal location 1q32.2-q41)
in the sample; and
comparing the expression of the genes, via the cDNA molecules, to the expression of said genes in reference samples of known human grade III breast cancer; and
identifying said sample as containing grade III breast cancer cells.
4 . The method of claim 1 wherein said assaying comprises preparing RNA from said sample.
5 . The method of claim 4 wherein said RNA is amplified by quantitative PCR to produce cDNA.
6 . The method of claim 4 wherein said RNA is amplified by reverse transcription PCR (RT-PCR) to produce cDNA.
7 . The method of claim 1 wherein said assaying comprises determining gene expression by an array.
8 . The method of claim 1 wherein said sample is a ductal lavage or fine needle aspirate sample.
9 . The method of claim 7 wherein said sample is microdissected to isolate one or more cells suspected of being breast cancer cells and said assaying is of RNA in said isolated cells.
10 . A method to determine the grade of breast cancer cells in a sample from a human subject, said method comprising
assaying said sample for gene expression that is decreased in a high proliferation index of grade III relative to grade I breast cancer, wherein said gene expression is assayed by synthesis of cDNA molecules.
11 . The method of claim 10 wherein said assaying comprises preparing RNA from said sample.
12 . The method of claim 11 wherein said RNA is amplified by quantitative PCR to produce cDNA molecules.
13 . The method of claim 11 wherein said RNA is amplified by reverse transcription PCR (RT-PCR) to produce cDNA molecules.
14 . The method of claim 10 wherein said assaying comprises determining gene expression by an array.
15 . The method of claim 10 wherein said sample is a ductal lavage or fine needle aspirate sample.
16 . The method of claim 15 wherein said sample is microdissected to isolate one or more cells suspected of being breast cancer cells and said assaying is of RNA in said isolated cells.