Detection of Cancer Cells in Body Fluids

Abstract

A method of detecting circulating melanoma or carcinoma cells in a subject. The method comprises obtaining a body fluid from a subject and detecting the expression of a panel of genes in the body fluid, wherein the expression of the panel of genes indicates the presence of circulating melanoma or carcinoma cells in the subject. Genes useful for detecting melanoma cells includes GalNAc-T, MAGE-A3, MART-1, PAX-3, and TRP-2; genes useful for detecting carcinoma cells include C-Met, MAGE-A3, Stanniocalcin-1, Stanniocalcin-2, mammaglobin, HSP27, GalNAc-T, CK20, and β-HCG. Also disclosed are kits containing agents for detecting the expression of these genes.

1 . A method of detecting circulating melanoma cells in a subject, comprising:
 obtaining a body fluid from a subject; and   detecting the expression of a panel of genes in the body fluid, the panel of genes including GalNAc-T and PAX-3, wherein the expression of the panel of genes indicates the presence of circulating melanoma cells in the subject.   
     
     
         2 . The method of  claim 1 , wherein the panel of genes further includes one or more additional genes selected from the group consisting of GalNAc-T, MAGE-A3, MART-1, PAX-3, TRP-2, MITF, and Tyrosinase. 
     
     
         3 - 16 . (canceled) 
     
     
         17 . A kit for implementing the method of  claim 1 , comprising a plurality of agents for detecting the expression of a panel of genes, wherein the panel of genes includes PAX-3 and one or more genes selected from the group consisting of GalNAc-T, MAGE-A3, MART-1, TRP-2, MITF, and Tyrosinase. 
     
     
         18 . The kit of  claim 17 , wherein the panel of genes includes PAX-3, MART-1, and MAGE-3; PAX-3, MART-1, and GalNAc-T; PAX-3, MAGE-3, and GalNAc-T; GalNAc-T, MAGE-A3, MART-1, and PAX-3; MART-1, GalNAc-T, MITF, and PAX-3; MART-1, TRP-2, GalNAc-T, and PAX-3; or Tyrosinase, MART-1, GalNAc-T, and PAX-3. 
     
     
         19 . The kit of  claim 17  wherein the plurality of agents for detecting the expression of a panel of genes includes at least three pairs of primers for respectively quantifying the expression levels of at least three genes selected from the group consisting of GalNAc-T, MAGE-A3, MART-1, PAX-3, TRP-2, MITF, and Tyrosinase; and enzymes and reagents for performing qRT. 
     
     
         20 . The kit of  claim 19 , comprising at least four pairs of primers for respectively quantifying the expression levels of at least four genes selected from the group consisting of GalNAc-T, MAGE-A3, MART-1, PAX-3, TRP-2, MITF, and Tyrosinase. 
     
     
         21 . The kit of  claim 20 , comprising at least four pairs of primers for respectively quantifying the expression levels of GalNAc-T, MAGE-A3, MART-1, and PAX-3. 
     
     
         22 - 24 . (canceled) 
     
     
         25 . A method of detecting circulating breast, gastric, pancreatic, or colon cancer cells in a subject, comprising:
 obtaining a body fluid from a subject; and   quantifying the expression levels of a panel of genes using qRT, the panel of genes including at least three genes selected from the group consisting of C-Met, MAGE-A3, Stanniocalcin-1, Stanniocalcin-2, mammaglobin, HSP27, GalNAc-T, CK20, and β-HCG, wherein the expression of the panel of genes indicates the presence of circulating breast, gastric, pancreatic, or colon cancer cells in the subject.   
     
     
         26 . The method of  claim 25 , wherein the body fluid is blood, bone marrow, cerebral spinal fluid, peritoneal fluid, or pleural fluid. 
     
     
         27 . The method of  claim 25 , wherein at least one of the panel of genes is expressed. 
     
     
         28 . The method of  claim 27 , wherein at least two of the panel of genes are expressed. 
     
     
         29 - 30 . (canceled) 
     
     
         31 . The method of  claim 25 , wherein the expression of the panel of genes indicates that the subject is suffering from or at risk for developing a subclinical systemic breast, gastric, pancreatic, or colon cancer. 
     
     
         32 . The method of  claim 25 , further comprising:
 quantifying the breast, gastric, pancreatic, or colon cancer status; assigning a clinical breast, gastric, pancreatic, or colon cancer stage to the subject;   predicting treatment response, breast, gastric, pancreatic, or colon cancer recurrence, or survival of the subject; monitoring breast, gastric, pancreatic, or colon cancer progression or treatment response;   selecting or randomizing a treatment regime; or a combination thereof in accordance with the expression of the panel of genes.   
     
     
         33 . The method of  claim 32 , wherein the treatment response, breast, gastric, pancreatic, or colon cancer recurrence, or survival of the subject is predicted for a period of at least three years following the removal of a primary tumor, SLND, or both. 
     
     
         34 . The method of  claim 25 , wherein the body fluid or a tissue sample from the subject is histopathologically negative for breast, gastric, pancreatic, or colon cancer cells. 
     
     
         35 . The method of  claim 34 , wherein the histopathology of the body fluid or tissue sample is determined by H&E or IHC staining. 
     
     
         36 . The method of  claim 25 , wherein the panel of genes includes at least four genes selected from the group consisting of C-Met, MAGE-A3, Stanniocalcin-1, Stanniocalcin-2, mammaglobin, HSP27, GalNAc-T, CK20, and β-HCG. 
     
     
         37 . The method of  claim 37 , wherein the panel of genes includes a first combination of C-Met, MAGE-A3, GalNAc-T, and CK20; a second combination of mammaglobin, C-Met, GalNAc-T, and β-HCG; a third combination of mammaglobin, β-HCG, HSP27, and C-Met; or a fourth combination of HSP27, CK20, Stanniocalcin-1, and MAGE-A3. 
     
     
         38 . (canceled) 
     
     
         39 . A kit, comprising a plurality of agents for detecting the expression of a panel of genes, wherein the panel of genes includes CK20, β-HCG and mammaglobin; GalNAc-T, mammaglobin, and β-HCG; mammaglobin, C-Met, GalNAc-T, and β-HCG; mammaglobin, β-HCG, HSP27, and C-Met; or HSP27, CK20, Stanniocalcin-1, and MAGE-A3. 
     
     
         40 . The kit of  claim 39  wherein the plurality of agents for detecting the expression of a panel of genes includes at least three pairs of primers for respectively quantifying the expression levels of at least three genes selected from the group consisting of C-Met, MAGE-A3, Stanniocalcin-1, Stanniocalcin-2, mammaglobin, HSP27, GalNAc-T, CK20, and β-HCG; and enzymes and reagents for performing qRT. 
     
     
         41 - 42 . (canceled)