Oncoprotein DEK as a tissue and urinary biomarker for bladder cancer

Antara Datta, Martin E Adelson, Yakov Mogilevkin, Eli Mordechai, Abraham A Sidi, Jason P Trama, Antara Datta, Martin E Adelson, Yakov Mogilevkin, Eli Mordechai, Abraham A Sidi, Jason P Trama

Abstract

Background: Bladder cancer is a significant healthcare problem in the United States of America with a high recurrence rate. Early detection of bladder cancer is essential for removing the tumor with preservation of the bladder, avoiding metastasis and hence improving prognosis and long-term survival. The objective of this study was to analyze the presence of DEK protein in voided urine of bladder cancer patients as a urine-based bladder cancer diagnostic test.

Methods: We examined the expression of DEK protein by western blot in 38 paired transitional cell carcinoma (TCC) bladder tumor tissues and adjacent normal tissue. The presence of DEK protein in voided urine was analyzed by western blot in 42 urine samples collected from patients with active TCC, other malignant urogenital disease and healthy individuals.

Results: The DEK protein is expressed in 33 of 38 bladder tumor tissues with no expression in adjacent normal tissue. Based on our sample size, DEK protein is expressed in 100% of tumors of low malignant potential, 92% of tumors of low grade and in 71% of tumors of high grade. Next, we analyzed 42 urine samples from patients with active TCC, other malignant urogenital disease, non-malignant urogenital disease and healthy individuals for DEK protein expression by western blot analysis. We are the first to show that the DEK protein is present in the urine of bladder cancer patients. Approximately 84% of TCC patient urine specimens were positive for urine DEK.

Conclusion: Based on our pilot study of 38 bladder tumor tissue and 42 urine samples from patients with active TCC, other malignant urogenital disease, non-malignant urogenital disease and healthy individuals; DEK protein is expressed in bladder tumor tissue and voided urine of bladder cancer patients. The presence of DEK protein in voided urine is potentially a suitable biomarker for bladder cancer and that the screening for the presence of DEK protein in urine can be explored as a noninvasive diagnostic test for bladder cancer.

Figures

Figure 1
Figure 1
DEK expression in cell lines. A. Whole cell lysates were prepared from bladder cancer cell lines (RT-4, 5637, T-24 and TCCSUP), Urothelial cell line (UroTSA) and undifferentiated and differentiated progenitor bladder epithelial cell lines. DEK protein was evaluated by western blot analysis using a DEK monoclonal antibody (BD Bioscience). β-actin was used as a loading control. B. Expression of DEK protein in whole cell (WC) lysate and respective conditioned media from 5637, 5637 expressing nonspecific shRNA (Ns shRNA), DEK shRNA or overexpressing DEK with a V5 tag (DEK-V5) by western blot assay using a monoclonal DEK antibody. Bottom panel represents a Coomassie stained gel to show that equal amounts of proteins were loaded on the gel.
Figure 2
Figure 2
DEK expression in bladder tumor tissue. Tissue lysates from bladder tumor tissue (T) or paired adjacent normal tissue (N) were analyzed by western blot using DEK monoclonal antibody (BD Biosciences). An established bladder SV-40 transformed urothelial cell line (UroTSA) lysate was used as a positive control. β-actin was used as a loading control. Sample MDL815 represents a biopsy confirmed inflamed tissue from bladder (no tumor present). Three representative western blots are shown.
Figure 3
Figure 3
DEK expression in urine. Thirty (30) ul of acetone precipitated urine protein lysates were analyzed by western blot using a polyclonal DEK antibody (Bethyl Laboratories). An established bladder SV-40 transformed urothelial cell line (UroTSA) lysate was used as a positive control. CAP indicates prostate cancer, RCC indicates renal cell carcinoma, H.TCC indicates history of transitional cell carcinoma and TCC indicates transitional cell carcinoma of the bladder. Voided urine sample, MDL 494* was collected from a suspected TCC patient but pathology report was inconclusive. Two representative western blots are shown.

References

    1. Society AC. American Cancer Society. Altanta; 2010. Cancer Facts and Figures 2010.
    1. Kaufman DS, Shipley WU, Feldman AS. Bladder cancer. Lancet. 2009;374:239–249. doi: 10.1016/S0140-6736(09)60491-8.
    1. Hurst RE. Does the biomarker search paradigm need re-booting? BMC Urol. 2009;9:1. doi: 10.1186/1471-2490-9-1.
    1. Van Tilborg AA, Bangma CH, Zwarthoff EC. Bladder cancer biomarkers and their role in surveillance and screening. Int J Urol. 2009;16:23–30. doi: 10.1111/j.1442-2042.2008.02174.x.
    1. Budman LI, Kassouf W, Steinberg JR. Biomarkers for detection and surveillance of bladder cancer. Can Urol Assoc J. 2008;2:212–221.
    1. Costa VL, Henrique R, Danielsen SA, Duarte-Pereira S, Eknaes M, Skotheim RI, Rodrigues A, Magalhaes JS, Oliveira J, Lothe RA, Three epigenetic biomarkers, GDF15, TMEFF2, and VIM, accurately predict bladder cancer from DNA-based analyses of urine samples. Clin Cancer Res. pp. 5842–5851.
    1. Jamshidian H, Kor K, Djalali M. Urine concentration of nuclear matrix protein 22 for diagnosis of transitional cell carcinoma of bladder. Urol J. 2008;5:243–247.
    1. von Lindern M, Breems D, van Baal S, Adriaansen H, Grosveld G. Characterization of the translocation breakpoint sequences of two DEK-CAN fusion genes present in t(6;9) acute myeloid leukemia and a SET-CAN fusion gene found in a case of acute undifferentiated leukemia. Genes Chromosomes Cancer. 1992;5:227–234. doi: 10.1002/gcc.2870050309.
    1. Wu Q, Hoffmann MJ, Hartmann FH, Schulz WA. Amplification and overexpression of the ID4 gene at 6p22.3 in bladder cancer. Mol Cancer. 2005;4:16. doi: 10.1186/1476-4598-4-16.
    1. Carro MS, Spiga FM, Quarto M, Di Ninni V, Volorio S, Alcalay M, Muller H. DEK Expression is controlled by E2F and deregulated in diverse tumor types. Cell Cycle. 2006;5:1202–1207. doi: 10.4161/cc.5.11.2801.
    1. Kroes RA, Jastrow A, McLone MG, Yamamoto H, Colley P, Kersey DS, Yong VW, Mkrdichian E, Cerullo L, Leestma J, Moskal JR. The identification of novel therapeutic targets for the treatment of malignant brain tumors. Cancer Lett. 2000;156:191–198. doi: 10.1016/S0304-3835(00)00462-6.
    1. Szer IS, Sierakowska H, Szer W. A novel autoantibody to the putative oncoprotein DEK in pauciarticular onset juvenile rheumatoid arthritis. J Rheumatol. 1994;21:2136–2142.
    1. Dong X, Michelis MA, Wang J, Bose R, DeLange T, Reeves WH. Autoantibodies to DEK oncoprotein in a patient with systemic lupus erythematosus and sarcoidosis. Arthritis Rheum. 1998;41:1505–1510. doi: 10.1002/1529-0131(199808)41:8<1505::AID-ART23>;2-N.
    1. Wise-Draper TM, Allen HV, Jones EE, Habash KB, Matsuo H, Wells SI. Apoptosis inhibition by the human DEK oncoprotein involves interference with p53 functions. Mol Cell Biol. 2006;26:7506–7519. doi: 10.1128/MCB.00430-06.
    1. Wise-Draper TM, Allen HV, Thobe MN, Jones EE, Habash KB, Munger K, Wells SI. The human DEK proto-oncogene is a senescence inhibitor and an upregulated target of high-risk human papillomavirus E7. J Virol. 2005;79:14309–14317. doi: 10.1128/JVI.79.22.14309-14317.2005.
    1. Wise-Draper TM, Mintz-Cole RA, Morris TA, Simpson DS, Wikenheiser-Brokamp KA, Currier MA, Cripe TP, Grosveld GC, Wells SI. Overexpression of the cellular DEK protein promotes epithelial transformation in vitro and in vivo. Cancer Res. 2009;69:1792–1799. doi: 10.1158/0008-5472.CAN-08-2304.
    1. Sanchez-Carbayo M, Socci ND, Lozano JJ, Li W, Charytonowicz E, Belbin TJ, Prystowsky MB, Ortiz AR, Childs G, Cordon-Cardo C. Gene discovery in bladder cancer progression using cDNA microarrays. Am J Pathol. 2003;163:505–516. doi: 10.1016/S0002-9440(10)63679-6.
    1. Mor-Vaknin N, Punturieri A, Sitwala K, Faulkner N, Legendre M, Khodadoust MS, Kappes F, Ruth JH, Koch A, Glass D. et al.The DEK nuclear autoantigen is a secreted chemotactic factor. Mol Cell Biol. 2006;26:9484–9496. doi: 10.1128/MCB.01030-06.
    1. Evans AJ, Gallie BL, Jewett MA, Pond GR, Vandezande K, Underwood J, Fradet Y, Lim G, Marrano P, Zielenska M, Squire JA. Defining a 0.5-mb region of genomic gain on chromosome 6p22 in bladder cancer by quantitative-multiplex polymerase chain reaction. Am J Pathol. 2004;164:285–293. doi: 10.1016/S0002-9440(10)63118-5.

Source: PubMed

赞助者和合作者

医疗条件

药物干预

3
订阅