Abstract
Although various samples, including tissue, cells, serum, and urine, from patients with renal cell carcinoma (RCC) have been analyzed, biomarkers with diagnostic value have yet to be identified. We used a proteomics approach to analyze cyst fluid in cases of cyst-associated RCC to identify accessible and abundant proteins that are overexpressed and/or secreted by RCC cells. Proteins in the cyst fluid were separated by reverse-phase high-performance liquid chromatography and agarose two-dimensional gel electrophoresis and were identified by tandem mass spectrometry. We conducted a National Center for Biotechnology Information search and a MEDLINE search to predict the function of these identified proteins and to select a tumor-marker candidate protein. Our search resulted in the identification and selection of the differentially regulated protein known as 14-3-3 protein beta/alpha, which was overexpressed in cyst fluid from cyst-associated RCC but has not been previously associated with RCC. We then measured its incidence through Western blotting of various normal and RCC samples (serum, urine, tissue, and cyst fluid). The expression levels of 14-3-3 protein beta/alpha were higher in urine samples from patients with RCC than in samples from healthy volunteers. Receiver operating characteristic (ROC) curve analyses were performed to assess this potential biomarker; these data (area under the ROC curve value was 0.8813) indicate a high degree of accuracy for this screening method. 14-3-3 Protein beta/alpha may be a diagnostically useful biomarker for early diagnosis of RCC.
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References
Amato RJ (2005) Renal cell carcinoma: review of novel single agent therapeutics and combination regimens. Ann Oncol 16:7–15
Hafez KS, Fergany AF, Novick AC (1999) Nephron sparing surgery for localized renal cell carcinoma: impact of tumor size on patient survival, tumor recurrence and TNM staging. J Urol 162:1930–1933
Patard JJ, Leray E, Cindolo L, Ficarra V, Rodriguez A, De La Taille A, Tostain J, Artibani W, Abbou CC, Guillé F, Chopin DK, Lobel B (2004) Multi-institutional validation of a symptom based classification for renal cell carcinoma. J Urol 172:858–862
Pantuck AJ, Zisman A, Belldegrun A (2001) The changing natural history of renal cell carcinoma. J Urol 166:1611–1623
Tunuguntla HS, Jorda M (2008) Diagnostic and prognostic molecular markers in renal cell carcinoma. J Urol 179:2096–2102
Seliger B, Dressler SP, Lichtenfels R, Kellner R (2007) Candidate biomarkers in renal cell carcinoma. Proteomics 24:4601–4612
Seliger B, Lichtenfels R, Atkins D, Bukur J, Halder T, Kersten M, Harder A, Ackermann A, Malenica B, Brenner W, Zobawa M, Lottspeich F (2005) Identification of fatty acid binding proteins as markers associated with the initiation and/or progression of renal cell carcinoma. Proteomics 10:2631–2640
Tolson J, Bogumil R, Brunst E, Beck H, Elsner R, Humeny A, Kratzin H, Deeg M, Kuczyk M, Mueller GA, Mueller CA, Flad T (2004) Serum protein profiling by SELDI mass spectrometry: detection of multiple variants of serum amyloid alpha in renal cancer patients. Lab Invest 84:845–856
Kim DS, Choi YP, Kang S, Gao MQ, Kim B, Park HR, Choi YD, Lim JB, Na HJ, Kim HK, Nam YP, Moon MH, Yun HR, Lee DH, Park WM, Cho NH (2008) Panel of candidate biomarkers for renal cell carcinoma. J Proteome Res 7:3710–3719
Sarkissian G, Fergelot P, Lamy PJ, Patard JJ, Culine S, Jouin P, Rioux-Leclercq N, Darbouret B (2008) Identification of pro-MMP-7 as a serum marker for renal cell carcinoma by use of proteomic analysis. Clin Chem 54:574–581
Gnarra JR, Lerman MI, Zbar B, Linehan WM (1995) Genetics of renal-cell carcinoma and evidence for a critical role for von Hippel-Lindau in renal tumorigenesis. Semin Oncol 22:3–8
Greene FL, Page DL, Fleming ID, Fritz A, Balch CM, Haller DG, Morrow M (2002) AJCC Cancer Staging Manual, 6th edn. Springer, New York
Kawashima Y, Fukuno T, Satoh M, Takahashi H, Matsui T, Tadakazu M, Yoshio K (2009) A simple and highly reproducible method for discovering potential disease markers in low abundance serum proteins. J Electrophoresis 53:13–18
Oh-Ishi M, Satoh M, Maeda T (2000) Preparative two-dimensional gel electrophoresis with agarose gels in the first dimension for high molecular mass proteins. Electrophoresis 21:1653–1669
Yates JR 3rd, Eng JK, McCormack AL, Schieltz D (1995) Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database. Anal Chem 67:1426–1436
Wulfkuhle JD, Paweletz CP, Steeg PS, Petricoin EF 3rd, Liotta L (2003) Proteomic approaches to the diagnosis, treatment, and monitoring of cancer. Adv Exp Med Biol 532:59–68
Aitken A (2006) 14-3-3 Proteins: a historic overview. Semin Cancer Biol 16:162–172
Rushworth LK, Hindley AD, O’Neill E, Kolch W (2006) Regulation and role of Raf-1/B-Raf heterodimerization. Mol Cell Biol 26:2262–2272
Sunayama J, Tsuruta F, Masuyama N, Gotoh Y (2005) JNK antagonizes Akt-mediated survival signals by phosphorylating 14-3-3. J Cell Biol 170:295–304
Tsuruta F, Sunayama J, Mori Y, Hattori S, Shimizu S, Tsujimoto Y, Yoshioka K, Masuyama N, Gotoh Y (2004) JNK promotes Bax translocation to mitochondria through phosphorylation of 14-3-3 proteins. EMBO J 23:1889–1999
Porter GW, Khuri FR, Fu H (2006) Dynamic 14-3-3/client protein interactions integrate survival and apoptotic pathways. Semin Cancer Biol 16:193–202
Mils V, Baldin V, Goubin F, Pinta I, Papin C, Waye M, Eychene A, Ducommun B (2000) Specific interaction between 14-3-3 isoforms and the human CDC25B phosphatase. Oncogene 19:1257–1265
Morrison DK (2009) The 14-3-3 proteins: integrators of diverse signaling cues that impact cell fate and cancer development. Trends Cell Biol 19:16–23
Moreira JM, Ohlsson G, Rank FE, Celis JE (2005) Down-regulation of the tumor suppressor protein 14-3-3sigma is a sporadic event in cancer of the breast. Mol Cell Proteomics 4:555–569
Moreira JM, Gromov P, Celis JE (2004) Expression of the tumor suppressor protein 14-3-3sigma is down-regulated in invasive transitional cell carcinomas of the urinary bladder undergoing epithelial-tomesenchymal transition. Mol Cell Proteomics 3:410–419
Lodygin D, Hermeking H (2005) The role of epigenetic inactivation of 14-3-3sigma in human cancer. Cell Res 15:237–246
Vogelzang NJ, Stadler WM (1998) Kidney cancer. Lancet 352:1691–1696
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This study was supported in part by the Ministry of Education, Culture, Sports, Science, and Technology of Japan (No. 22591776).
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Minamida, S., Iwamura, M., Kodera, Y. et al. 14-3-3 Protein beta/alpha as a urinary biomarker for renal cell carcinoma: proteomic analysis of cyst fluid. Anal Bioanal Chem 401, 245–252 (2011). https://doi.org/10.1007/s00216-011-5057-5
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DOI: https://doi.org/10.1007/s00216-011-5057-5