Genes and polypeptides relating to hepatocellular or colorectal carcinoma

Abstract

The present application provides novel human genes WDRPUH and KRZFPUH and PPIL1 whose expression is markedly elevated in a great majority of HCCs and colorectal cancers, respectively, compared to corresponding non-cancerous tissues, as well as novel human gene APCDD1 whose expression is elevated in primary colon cancers and down-regulated in response to the transduction of wild-type APC1 into colon-cancer cells. The genes and polypeptides encoded by the genes can be used, for example, in the diagnosis of a cell proliferative disease, and as target molecules for developing drugs against the disease.

1 . An A substantially pure polypeptide selected from the group consisting of: 
 (a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of any one of SEQ ID NO: 2, 4, 6, or 8.    
     
     
         2 . An isolated polynucleotide encoding the polypeptide of  claim 1 .  
     
     
         3 . A vector comprising the polynucleotide of  claim 2 .  
     
     
         4 . A host cell harboring the polynucleotide of  claim 2  or the vector of  claim 3 .  
     
     
         5 . A method for producing the polypeptide of  claim 1 , said method comprising the steps of: 
 (a) culturing a host cell comprising a polynucleotide encoding the polypeptide;    (b) allowing the host cell to express the polypeptide; and    (c) collecting the expressed polypeptide.    
     
     
         6 . An antibody binding to the polypeptide of  claim 1 .  
     
     
         7 . A polynucleotide that is complementary to the polynucleotide of  claim 2  or to the complementary strand thereof and that comprises at least 15 nucleotides.  
     
     
         8 . An antisense polynucleotide or small interfering RNA against the polynucleotide of  claim 2 .  
     
     
         9 . The antisense polynucleotide of  claim 8 , which is selected from the group consisting of nucleotides comprising the nucleotide sequence of SEQ ID NO: 16, 37, 44, and 89.  
     
     
         10 . The small interfering RNA of  claim 8 , wherein the sense strand thereof is selected from the group consisting of nucleotides comprising the nucleotide sequence of SEQ ID NOs: 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, and 103.  
     
     
         11 . A method for diagnosing a cell proliferative disease, said method comprising the steps of: 
 (a) detecting the expression level of the gene encoding the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in biological sample of specimen; and    (b) relating an elevation of the expression level to the disease.    
     
     
         12 . The method of  claim 11 , wherein the expression level is detected by any one method select from group consisting of: 
 (a) detecting the mRNA encoding the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;    (b) detecting the protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and    (c) detecting the biological activity of the protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8.    
     
     
         13 . A method of screening for a compound for treating a cell proliferative disease, said method comprising the steps of: 
 (a) contacting a test compound with a polypeptide selected from the group consisting of: 
 (1) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;  
 (2) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and  
 (3) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;  
   (b) detecting the binding activity between the polypeptide and the test compound; and    (c) selecting a compound that binds to the polypeptide.    
     
     
         14 . A method of screening for a compound for treating a cell proliferative disease, said method comprising the steps of: 
 (a) contacting a candidate compound with a cell expressing one or more polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7; and    (b) selecting a compound that reduces the expression level of one or more polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7 in comparison with the expression level detected in the absence of the test compound.    
     
     
         15 . A method of screening for a compound for treating a cell proliferative disease, said method comprising the steps of: 
 (a) contacting a test compound with a polypeptide selected from the group consisting of: 
 (1) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;  
 (2) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and  
 (3) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;  
   (b) detecting the biological activity of the polypeptide of step (a); and    (c) selecting a compound that suppresses the biological activity of the polypeptide in comparison with the biological activity detected in the absence of the test compound.    
     
     
         16 . The method of  claim 15 , wherein the biological activity is cell-proliferating activity.  
     
     
         17 . A method of screening for a compound for treating a cell proliferative disease, said method comprising the steps of: 
 (a) contacting a candidate compound with a cell into which a vector comprising the transcriptional regulatory region of one or more marker genes and a reporter gene that is expressed under the control of the transcriptional regulatory region has been introduced, wherein the one or more marker genes comprise any one of nucleotide sequence selected from the group consisting of SEQ ID:NO 1, 3, 5, or 7,    (b) measuring the activity of said reporter gene; and    (c) selecting a compound that reduces the expression level of said reporter gene as compared to a control.    
     
     
         18 . A method of screening for a compound for treating a cell proliferative disease, said method comprising the steps of: 
 (a) constructing a vector comprising the two Tcf/LEF binding motifs of APCDD1 upstream of a reporter gene;    (b) transforming a cell with the vector of step (a);    (c) contacting a test compound with the cell of step (b) in the existence of β-catenin/Tcf 4 complex;    (d) detecting the expression of the reporter gene; and    (e) electing the test compound that suppresses the expression of the reporter gene compared to that in the absence of the test compound.    
     
     
         19 . A method of screening for a compound for treating a cell proliferative disease, said method comprising the steps of: 
 (a) contacting a polypeptide selected from the group consisting of: 
 (1) a polypeptide comprising the amino acid sequence of SEQ ID NO: 6;  
 (2) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 6 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of the amino acid sequence of SEQ ID NO: 6; and  
   (3) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 5, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6;    with stathmin or SNW1 in the existence of a test compound; 
 (b) detecting the binding between the polypeptide and stathmin or SNW1; and  
 (c) selecting the test compound that inhibits the binding between the polypeptide and stathmin or SNW1.  
   
     
     
         20 . A method of any one of  claim 11  to  19 , wherein the cell-proliferative disease is cancer.  
     
     
         21 . A composition for treating a cell proliferative disease, said composition comprising a pharmaceutically effective amount of an antisense polynucleotide or small interfering RNA against a polynucleotide encoding a polypeptide selected from the group consisting of: 
 (a) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8.    
     
     
         22 . A composition for treating a cell proliferative disease, said composition comprising a pharmaceutically effective amount of an antibody against a polypeptide selected from the group consisting of: 
 (a) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8.    
     
     
         23 . A composition for treating a cell proliferative disease, said composition comprising a pharmaceutically effective amount of the compound selected by the method of any one of  claims 13  to  19 .  
     
     
         24 . The composition of any one of claims  21  and  22 , wherein the cell proliferative disease is cancer.  
     
     
         25 . A method for treating a cell proliferative disease, said method comprising the step of administering a pharmaceutically effective amount of an antisense polynucleotide or small interfering RNA against a polynucleotide encoding a polypeptide selected from the group consisting of: 
 (a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8.    
     
     
         26 . A method for treating a cell proliferative disease, said method comprising the step of administering a pharmaceutically effective amount of an antibody against a polypeptide selected from the group consisting of: 
 (a) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8;    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8; and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8.    
     
     
         27 . A method for treating a cell proliferative disease, said method comprising the step of administering a pharmaceutically effective amount of a compound selected by the method of any one of  claims 13  to  19 .  
     
     
         28 . The method of any one of claims  25  and  26 , wherein the cell proliferative disease is cancer.  
     
     
         29 . A method for treating or preventing a cancer, said method comprising the step of administering a pharmaceutically effective amount of a polypeptide selected from the group consisting of (a)-(c), or a polynucleotide encoding the polypeptide: 
 (a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof;    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof; and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof.    
     
     
         30 . A method for inducing an anti tumor immunity, said method comprising the step of contacting a polypeptide selected from the group consisting of (a)-(c) with antigen presenting cells, or introducing a polynucleotide encoding the polypeptide or a vector comprising the polynucleotide to antigen presenting cells: 
 (a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof;    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof, and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof.    
     
     
         31 . The method for inducing an anti tumor immunity of  claim 30 , wherein the method further comprises the step of administering the antigen presenting cells to a subject.  
     
     
         32 . A pharmaceutical composition for treating or preventing a cancer, said composition comprising a pharmaceutically effective amount of polypeptide selected from the group of (a)-(c), or a polynucleotide encoding the polypeptide: 
 (a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof,    (b) a polypeptide that comprises the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8 in which one or more amino acids are substituted, deleted, inserted, and/or added and that has a biological activity equivalent to a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof; and    (c) a polypeptide encoded by a polynucleotide that hybridizes under stringent conditions to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1, 3, 5, or 7, wherein the polypeptide has a biological activity equivalent to a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, or 8, or a fragment thereof.    
     
     
         33 . The pharmaceutical composition of  claim 32 , wherein the polynucleotide is incorporated in expression vector.  
     
     
         34 . The composition of  claim 23 , wherein the cell proliferative disease is cancer.  
     
     
         35 . The method of  claim 27 , wherein the cell proliferative disease is cancer.