Method of assessing cancerous conditions and reagent for detecting gene product to be used in the method

Abstract

It is intended to provide a method of assessing cancerous conditions, which is useful in diagnosing cancer in a therapy or a preventive therapy for cancer diseases or a maintenance therapy therefor, and a reagent for detecting a gene product to be used in the above method. The cancerous conditions of a human-origin specimen is assessed by using, as an indication, the content of a gene product, which shows an increase or a decrease in the expression thereof due to an unusual alternative splicing, from gene products obtained from human PTCH1 gene contained in the human-origin specimen. To measure the content, use is preferably made of, concerning at least one base sequence selected from the group consisting of (a) the base sequences represented by SEQ ID NOS:1 to 4 and (b) base sequences derived from any of the base sequences represented by SEQ ID NOS:1 to 4 by deletion, substitution and/or insertion of one to several bases and having an activity of being hybridized with a chain complementary to the corresponding base sequence selected from the base sequences represented by SEQ ID NOS:1 to 4 under stringent conditions, a probe sequence-specifically binding to the complementary chain of the above-described base sequence or a chemical modification thereof.

1 . A method of determining cancerous conditions of a human-derived sample, comprising determining cancerous conditions of the human-derived sample by using an abundance of a gene product of human PTCH1 gene contained in the sample as an index, the expression of the gene product from gene products of said human PTCH1 gene being varied due to the modulation in a alternative splicing. 
     
     
         2 . A method of determining cancerous conditions of a human-derived sample, comprising the steps of:
 (1) measuring an abundance of at least one transcription product from transcription products of human PTCH1 gene contained in the human-derived sample, wherein the transcription product of which the abundance is measured contains, in an RNA sequence thereof, at least one base sequence selected from the group consisting of (a) base sequences described in SEQ ID Nos. 1 to 4, and (b) base sequences having deletion, substitution, and/or Insertion of one to several bases in the base sequences described in SEQ ID Nos. 1 to 4 and having ability to hybridize respectively with corresponding complementary strands of the base sequences described in SEQ ID Nos. 1 to 4 under stringent conditions; and   (2) determining cancerous conditions of the human-derived sample by using the measured abundance of the transcription product in the human-derived sample as an index.   
     
     
         3 . The method of determining cancerous conditions according to  claim 2 , further comprising the step of measuring an abundance of at least one transcription product from transcription products of human PTCH1 gene contained in a control sample which is obtained from a normal human living body tissue,
 wherein the transcription product of which the abundance in the control sample is measured contains, in an RNA sequence thereof, at least one base sequence selected from the group consisting of (a) base sequences described in SEQ ID Nos. 1 to 4, and (b) base sequences having deletion, substitution, and/or insertion of one to several bases in the base sequences described in SEQ ID Nos. 1 to 4 and having ability to hybridize respectively with corresponding complementary strands of the base sequences described in SEQ ID Nos. 1 to 4 under stringent conditions, and   wherein the step of determining cancerous conditions of the human-derived sample is performed by comparing the measured abundance of transcription product in the human-derived sample with the measured abundance of the transcription product in the control sample.   
     
     
         4 . The method of determining cancerous conditions according to  claim 2  or  3 , wherein the abundance of the transcription product is measured by a method in which the transcription product is detected by using a probe or chemical modulation thereof that binds sequence-specifically to the complementary strand of at least one base sequence selected from the group consisting of (a) bases sequences described in SEQ ID Nos. 1 to 4, and (b) base sequences having deletion, substitution, and/or insertion of one to several bases in the base sequences described in SEQ ID Nos. 1 to 4 and having ability to hybridize respectively with corresponding complementary strands of the base sequences described in SEQ ID Nos. 1 to 4 under stringent conditions. 
     
     
         5 . The method of determining cancerous conditions according to  claim 4 , wherein the probe or chemical modulation thereof includes a polynucleotide, a polyribonucleotide, chemical modulations thereof, and a polynucleotide, polyribonucleotide or chemical modulations thereof having a base sequence selected from those sequences having substitution, insertion, and/or deletion of one or more bases such that sequence-specific binding ability to the complementary strand of nucleic acid is not impaired. 
     
     
         6 . A method of determining cancerous conditions of a human-derived sample, comprising the steps of:
 (1) measuring an abundance of a translation product from at least one transcription product from transcription products of human PTCH1 gene contained in the human-derived sample, wherein the transcription product from which the abundance of the translation product is measured contains, in an RNA sequence thereof, at least one base sequence selected from the group consisting of (a) base sequences described in SEQ ID Nos. 1 to 4, and (b) base sequences having deletion, substitution, and/or insertion of one to several bases in the base sequences described in SEQ ID Nos. 1 to 4 and having ability to hybridize respectively with corresponding complementary strands of the base sequences described in SEQ ID Nos. 1 to 4 under stringent conditions; and   (2) determining cancerous conditions of the human-derived sample by using the measured abundance of the translation product in the human-derived sample as an index.   
     
     
         7 . The method of determining cancerous conditions according to  claim 6 , further comprising the step of measuring an abundance of a translation product from at least one transcription product from transcript ion products of human PTCH1 gene contained in a control sample which is obtained from a normal human living body tissue,
 wherein the transcription product from which the abundance of the translation product in the control sample measured contains, in an RNA sequence thereof, at least one base sequence selected from the group consisting of (a) base sequences described in SEQ ID Nos. 1 to 4, and (b) base sequences having deletion, substitution, and/or insertion of one to several bases in the base sequences described in SEQ ID Nos. 1 to 4 and having ability to hybridize respectively with corresponding complementary strands of the base sequences described in SEQ ID Nos. 1 to 4 under stringent conditions, and   wherein the step of determining cancerous conditions of the human-derived sample is performed by comparing the measured abundance of the translation product in the human-derived sample with the measured abundance of the translation product in the control sample.   
     
     
         8 . The method of determining cancerous conditions according to  claim 3  or  7 , wherein the human-derived sample and the control sample obtained from the normal human living body tissue are samples obtained from the same subject. 
     
     
         9 . The method of determining cancerous conditions according to any one of  claims 1  to  8 , wherein the human-derived sample is a specimen obtained from a human. 
     
     
         10 . The method of determining cancerous conditions according to any one of  claims 1  to  9 , wherein the human-derived sample is a human living body tissue. 
     
     
         11 . The method of determining cancerous conditions according to any one of  claims 1  to  10 , wherein the human-derived sample is a sample extracted from at least one organ selected from the group consisting of colon, leukocyte, brain, and lung. 
     
     
         12 . The method of determining cancerous conditions according to any one of  claims 2  to  5 , wherein the method of measuring the abundance of the transcription product is any one of the measuring methods selected from the group consisting of a micro-array method, an RT-PCR method, a real time PCR method, a subtraction method, a differential display method, a differential hybridization method, or a cross hybridization method. 
     
     
         13 . The method of determining cancerous conditions according to  claim 6  to  7 , wherein the method of measuring the abundance of the translation product form the transcription product is a Western blotting method or an ELISA method. 
     
     
         14 . A reagent for detecting gene product, the reagent comprising a probe or chemical modulation that specifically binds to a gene product of human PTCH1 gene, the expression of the gene product from gene products of the human PTCH1 gene being varied due to the unusual alternative splicing, so that the reagent can be use for measuring the abundance of the gene product contained in a human-derived sample. 
     
     
         15 . The reagent for detecting gene product according to  claim 14 , wherein the gene product is a transcription product of human PTCH1 gene contained in the human-derived sample, and wherein the probe or chemical modulation thereof. Is a probe or chemical modulation that sequence-specifically binds to the complementary strand of nucleic acid of the transcription product whose expression is varied. 
     
     
         16 . The reagent for detecting gene product according to  claim 14 , wherein the gene product is a translation product from a transcription product of human PTCH1 gene contained in the human-derived sample, and wherein the probe or chemical modulation thereof is an antibody or chemical modulation thereof that specifically binds to the translation product whose expression is varied. 
     
     
         17 . The reagent for detecting gene product according to  claim 14  or  15 , wherein the probe or chemical modulation thereof includes a probe or chemical modulation thereof that binds sequence-specifically to the sequence selected from the group consisting of (a) base sequences described in SEQ ID Nos. 1 to 4, and (b) base sequences having deletion, substitution, and/or insertion of one to several bases in the base sequences described in SEQ ID Nos. 1 to 4 and having ability to hybridize respectively with corresponding complementary strands of the base sequences described in SEQ ID Nos. 1 to 4 under stringent conditions. 
     
     
         18 . The reagent for detecting gene product according to  claim 17 , wherein the probe or chemical modulation thereof. Includes a polynucleotide, a polyribonucleotide, chemical modulations thereof, and a polynucleotide, polyribonucleotide or chemical modulations thereof having a base sequence selected from those sequences having substitution, insertion, and/or deletion of one or more bases such that sequence-specific binding ability to the complementary strand of nucleic acid is not impaired. 
     
     
         19 . The reagent for detecting gene product according to  claim 18 , wherein the probe or chemical modulation thereof includes 15- to 30-base long oligonucleotide.