Diagnosis kit and chip for bladder cancer using bladder cancer specific methylation marker gene

Abstract

The present invention relates to a kit and nucleic acid chip for diagnosing bladder cancer using a bladder cancer-specific marker gene. More particularly, the invention relates to a kit and nucleic acid chip for diagnosing bladder cancer, which can detect the promoter methylation of a bladder cancer-specific gene, the promoter or exon region of which is methylated specifically in transformed cells of bladder cancer. The use of the diagnostic kit or nucleic acid chip of the invention enables diagnosis of bladder cancer at an early stage of transformation, thus enabling early diagnosis of bladder cancer, and can diagnose bladder cancer in a more accurate and rapid manner compared to a conventional method.

1 . A kit for diagnosing bladder cancer, which comprises the methylated promoter of a bladder cancer marker gene selected from the group consisting of:
 (1) CDX2 (NM — 001265)—caudal type homeobox transcription factor 2;   (2) CYP1B1 (NM — 000104)—cytochrome P450, family 1, subfamily B, polypeptide 1;   (3) VSX1 (NM — 199425)—visual system homeobox 1 homolog, CHX10-like (zebrafish);   (4) HOXA11 (NM — 005523)—homeobox A11;   (5) T (NM — 003181)—T, brachyury homolog (mouse);   (6) TBX5 (NM — 080717)—T-box 5;   (7) PENK (NM — 006211)—proenkephalin;   (8) PAQR9 (NM — 198504)—progestin and adipoQ receptor family member IV;   (9) LHX2 (NM — 004789)—LIM Homeobox 2; and   (10) SIM2 (U80456)—single-minded homolog 2 ( Drosophila ).   
     
     
         2 . The kit for diagnosing bladder cancer according to  claim 1 , wherein the promoter or exon region contains at least one methylated CpG dinucleotide. 
     
     
         3 . The kit for diagnosing bladder cancer according to  claim 1 , wherein the promoter or exon region is any one of DNA sequences represented in SEQ ID NO: 31 to SEQ ID NO: 40. 
     
     
         4 . A nucleic acid chip for diagnosing bladder cancer, which comprises a probe capable of hybridizing with a fragment containing the CpG island of the promoter of the bladder cancer marker gene selected from the group consisting of:
 (1) CDX2 (NM — 001265)—caudal type homeobox transcription factor 2;   (2) CYP1B1 (NM — 000104)—cytochrome P450, family 1, subfamily B, polypeptide 1;   (3) VSX1 (NM — 199425)—visual system homeobox 1 homolog, CHX10-like (zebrafish);   (4) HOXA11 (NM — 005523)—homeobox A11;   (5) T (NM — 003181)—T, brachyury homolog (mouse);   (6) TBX5 (NM — 080717)—T-box 5;   (7) PENK (NM — 006211)—proenkephalin;   (8) PAQR9 (NM — 198504)—progestin and adipoQ receptor family member IV;   (9) LHX2 (NM — 004789)—LIM Homeobox 2; and   (10) SIM2 (U80456)—single-minded homolog 2 ( Drosophila ).   
     
     
         5 . The nucleic acid chip for diagnosing bladder cancer according to  claim 4 , wherein the promoter is any one of DNA sequences represented in SEQ ID NO: 31 to SEQ ID NO: 40. 
     
     
         6 . A method for detecting the methylation of the promoter or exon region of a clinical sample-originated gene selected from the group consisting of CDX2, CYP1B1, VSX1, HOXA11, T, TBX5, PENK, PAQR9, LHX2 and SIM2. 
     
     
         7 . The method for detecting the methylation according to  claim 6 , wherein the method for measuring methylation is selected from the group consisting of PCR, methylation specific PCR, real-time methylation specific PCR, PCR using a methylated DNA-specific binding protein, quantitative PCR, pyrosequencing and bisulfite sequencing. 
     
     
         8 . The method for detecting the methylation according to  claim 6 , wherein the clinical sample is tissue, cell, blood or urine originated from patients suspected of cancer or subjects to be diagnosed.