Method of identifying cancer biomarkers and cancer progression

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase

Reexamination Certificate

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C435S215000

Reexamination Certificate

active

07662582

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.

REFERENCES:
Hasui et al. The Content of Urokinase-Type Plasminogen Activator and Tumor Recurrence in Superficial Bladder Cancer; The Journal of Urology, vol. 151 (1994) pp. 16-20.
Nielsen, L.S., et al. Enzyme-Linked Immunosorbent Assay for Human Urokinase-Type Plasminogen Activation and its Proenzyme Using a Combination of Monoclonal and Polyclonal Antibodies; Journal of Immunoassay, vol. 7, No. 3 (1986) pp. 209-228.
Kielberg et al. Proenzyme to Urokinase-Type Plasminogen Activator in the Mouse in Vivo; FEBS Letters, vol. 182, No. 2 (1985) pp. 441-445.
Chiao-Yun Lin, et al, Searching Cell-secreted proteomes for potential urinary bladder tumor markers, Wiley Interscience, www.interscience.wiley.com, vol. 6 Issue 15, Jul. 3, 2006.

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