Methods and nucleic acids for the analysis of gene expression associated with the development of prostate cell proliferative disorders

Abstract

The invention provides methods, nucleic acids and kits for detecting prostate cell proliferative disorders. 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 detecting prostate cell proliferative disorders in a subject comprising determining the expression levels of at least one gene selected from the group consisting of RASSF2A; TFAP2E; HIST1H4K and GSTPi in a biological sample isolated from said subject wherein underexpression and/or CpG methylation is indicative of the presence of said disorder. 
     
     
         2 . The method according to  claim 1  comprising determining the expression levels of the gene RASSF2A and at least one gene selected from the group consisting of TFAP2E; HIST1H4K and GSTPi. 
     
     
         3 . The method according to  claim 1  comprising determining the expression levels of the gene TFAP2E and at least one gene selected from the group consisting of RASSF2A; HIST1H4K and GSTPi. 
     
     
         4 . The method according to  claim 1  comprising determining the expression levels of the gene HIST1H4K and at least one gene selected from the group consisting of RASSF2A; TFAP2E and GSTPi. 
     
     
         5 . The method according to any of  claims 1  to  4  wherein said expression is determined by detecting the presence or absence of CpG methylation within said gene, wherein the presence of methylation indicates the presence of a carcinoma. 
     
     
         6 . The method according to  claim 5  wherein said CpG is located within a sequence that comprises, or hybridizes under stringent conditions to a sequence of at least 16 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4. 
     
     
         7 . A method according to any of  claims 1  to  6 , comprising contacting genomic DNA isolated from a biological sample obtained from said subject with at least one reagent, or series of reagents that distinguishes between methylated and non-methylated CpG dinucleotides within at least one target region of the genomic DNA, wherein the target region comprises, or hybridizes under stringent conditions to a sequence of at least 16 contiguous nucleotides of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4 respectively, wherein said contiguous nucleotides comprise at least one CpG dinucleotide sequence. 
     
     
         8 . The method of any of  claims 1  to  7 , wherein the biological sample obtained from the subject is selected from the group comprising cell lines, histological slides, biopsies, paraffin-embedded tissue, body fluids, ejaculate, ejaculate, urine, blood plasma, blood serum, whole blood, isolated blood cells, cells isolated from the blood and combinations thereof. 
     
     
         9 . A kit suitable for performing the method of any of  claims 1  to  8  comprising (a) a bisulfite reagent; (b) a container suitable for containing the said bisulfite reagent and the biological sample of the patient; (c) at least one set of oligonucleotides containing two oligonucleotides whose sequences in each case are identical, are complementary, or hybridize under stringent or highly stringent conditions to a 9 or more preferably 18 base long segment of a sequence selected from SEQ ID NO: 5 to SEQ ID NO: 20. 
     
     
         10 . A kit suitable for performing the method of any of  claims 1  to  9  comprising (a) a methylation sensitive restriction enzyme reagent; (b) a container suitable for containing the said reagent and the biological sample of the patient; (c) at least one set of oligonucleotides one or a plurality of nucleic acids or peptide nucleic acids which are identical, are complementary, or hybridize under stringent or highly stringent conditions to an at least 9 base long segment of a sequence selected from the group consisting of SEQ ID NO: 1 to SEQ ID NO: 4; and optionally (d) instructions for use and interpretation of the kit results.