Method of identifying cancer biomarkers and cancer progression

Abstract

An efficient method for identifying important cancer biomarkers and identifying progression of bladder cancer using pro-u-PA as a clinical tool is provided. Searching for biomarkers critical for bladder carcinoma diagnosis and prognosis, secreted proteomes of highly malignant U1 and pre-malignant U4 cell lines are first analyzed. Proteins in the cultured media of the U1 and U4 cell-lines were systematically examined by SDS-PAGE combined with MALDI-TOF mass spectrometry. Expression of pro-u-plasminogen activator (pro-u-PA) was confirmed by Western blot analysis and further evaluated. A statistically significant relationship between the low level and absence of pro-u-PA in urine with high stages and grades of the tumor samples was established. Constitutive expression of Ras dominant negative protein led to increased expression of pro-u-PA in cultured media, indicating the loss of pro-u-PA is associated with oncogenic transformation. The loss of pro-u-PA in urine has been identified as a marker of more advanced bladder carcinoma.

1 . A method for identifying important cancer biomarkers comprising:
 analyzing secreted proteomes of highly malignant U1 and pre-malignant U4 cell lines;   examining proteins in cultured media of the U1 and U4 cell-lines by SDS-PAGE combined with MALDI-TOF mass spectrometry; and   confirming an expression by Western blot analysis.   
   
   
       2 . The method for identifying important cancer biomarkers of  claim 1 , where the expression comprises an expression of pro-u-plasminogen activator. 
   
   
       3 . The method for identifying important cancer biomarkers of  claim 1 , where a level of pro-u-plasminogen identifies cancer progress. 
   
   
       4 . The method for identifying important cancer biomarkers of  claim 1 , where a low level and absence of pro-u-plasminogen activator indicates high stage and grade of a tumor. 
   
   
       5 . The method of identifying important cancer biomarkers of  claim 1 , where levels of heat shock protein 70, M2-type pyrurate kinase, glyceraldehydes-3-phosphate dehyrogenase, lactate dehydrogenase, phosphoglycerate kinase 1, and triosephosphate isomerase 1 in the cultured media identifies cancer progress.