PITX2 Polynucleotide, Polypeptide and Methods of Use Therefor

Abstract

A novel T-type calcium channel (CACNA1G) is provided, as are polynucleotides encoding the same. CACNA1G has been implicated in cellular proliferative disorders. More specifically, it has been observed that the methylation state of specific regions within CpG islands associated with the CACNA1G gene correlates with a number of cancerous phenotypes involving a variety of tissue and cell types. Also provided are methods for detecting cellular proliferative disorders by determining the methylation state of genes or regulatory regions associated therewith, including CACNA 1 G, as well as kits containing reagents for performing invention methods.

1 . A method for detecting leukemia, prostate cancer, or breast cancer in a human subject comprising:
 a) contacting a nucleic acid-containing specimen from the subject with an agent that provides a determination of the methylation state of a 5′ CpG rich region of a PITX2 gene, wherein the region consists of the sequence set forth in SEQ ID NO:115; and   b) identifying aberrant methylation of regions of the gene or regulatory region, wherein aberrant methylation comprises hypermethylation when compared to the same regions of the gene or associated regulatory region in a human subject not having said cancer, thereby detecting leukemia, prostate cancer, or breast cancer in the human subject.   
     
     
         2 . The method of  claim 1 , wherein the regions of said gene are contained within CpG rich regions. 
     
     
         3 . The method of  claim 1 , wherein aberrant methylation comprises hypermethylation when compared to the same regions of the gene or associated regulatory regions in a subject not having the cellular proliferative disorder. 
     
     
         4 . The method of  claim 1 , wherein the agent is a pair of primers that hybridize with a target sequence in the 5′ CpG rich region of the PITX2 gene, wherein the region consists of the sequence set forth in SEQ ID NO:115. 
     
     
         5 . The method of  claim 4 , wherein the primers hybridize with a target polynucleotide sequence having the sequence selected from the group consisting of SEQ ID NO:55-103 and SEQ ID NO:104. 
     
     
         6 . The method of  claim 5 , wherein the primers are in consecutive pairs selected from the group consisting of SEQ ID NO:23 and SEQ ID NO:24. 
     
     
         7 . The method of  claim 1 , wherein the nucleic acid-containing specimen comprises a tissue selected from the group consisting of brain, colon, urogenital, lung, renal, prostate, pancreas, liver, esophagus, stomach, hematopoietic, breast, thymus, testis, ovarian, and uterine. 
     
     
         8 . The method of  claim 1 , wherein the nucleic acid-containing specimen is selected from the group consisting of serum, urine, saliva, blood, cerebrospinal fluid, pleural fluid, ascites fluid, sputum, stool, and biopsy sample. 
     
     
         9 . The method of  claim 1 , wherein said cancer is selected from the group consisting of low grade astrocytoma, anaplastic astrocytoma, glioblastoma, medulloblastoma, gastric cancer, colorectal cancer, colorectal adenoma, acute myelogenous leukemia, lung cancer, renal cancer, leukemia, breast cancer, prostate cancer, endometrial cancer and neuroblastoma. 
     
     
         10 . The method of  claim 7 , wherein the specimen comprises breast, prostate or hematopoietic tissue. 
     
     
         11 . The method of  claim 9 , wherein the cancer is breast cancer, prostate cancer or leukemia. 
     
     
         12 . The method of  claim 1 , wherein the detecting is for diagnosis of leukemia, prostate cancer, or breast cancer. 
     
     
         13 . The method of  claim 1 , wherein the detecting is for prognosis of leukemia, prostate cancer, or breast cancer.