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01-06-2011 | Research

Implication of RAF and RKIP Genes in Urinary Bladder Cancer

Authors: Apostolos Zaravinos, Maria Chatziioannou, George I. Lambrou, Ioannis Boulalas, Dimitris Delakas, Demetrios A. Spandidos

Published in: Pathology & Oncology Research | Issue 2/2011

Abstract

RKIP has been shown to regulate the RAS-RAF-MEK-ERK kinase cascade acting as modulator of apoptosis and metastasis in prostate cancer. Our goal was to examine the expression of the RAF (A-RAF, B-RAF and RAF-1) and RKIP genes in urinary bladder cancer. Microarray analysis and qPCR was employed to investigate the expression of RAF and RKIP, in 30 patients with transitional cell carcinoma (TCC) of the urinary bladder vs. the corresponding levels of adjacent normal tissue. Computational analysis was also performed on Gene Expression Omnibus (GEO) datasets, to unravel differences in the expression of RAF or RKIP between tumor and control samples, and between superficial and muscle invasive tumors. Microarray analysis revealed >2-fold expression of BRAF and RKIP in T2, T3, grade III tumors vs. controls. B-RAF over-expression was verified by qPCR in pT1, grade III tumors vs. their normal counterparts (p = 0.016). qPCR revealed a significant RKIP reduction in TCC vs. normal tissue (p = 0.002 and p < 0.001 for T1, grade II and Ta-T1, grade III, respectively); All RAF genes were positively correlated among each other (A-RAF/B-RAF, p = 0.003; A-RAF/RAF-1, p < 0.001; B-RAF/RAF-1, p = 0.050), whereas B-RAF was negatively correlated with RKIP in TCC (p = 0.050). Further computational analysis revealed different expression profiles for the genes of interest, among muscle invasive carcinomas, superficial TCCs, cystectomy specimens and normal tissue. The reduced RKIP mRNA levels in TCC and the elevated levels of B-RAF in pT1, grade III tumors vs. normal tissue, corroborate that these genes are involved in the pathogenesis of urinary bladder cancer.

Jemal A, Siegel R, Ward E et al (2008) Cancer statistics. CA Cancer J Clin 58:71–96PubMedCrossRef

Lopez-Beltran A, Montironi R (2004) Non-invasive urothelial neoplasms: according to the most recent WHO classification. Eur Urol 46:170–176PubMedCrossRef

O’Neill E, Kolch W (2004) Conferring specificity on the ubiquitous Raf/MEK signalling pathway. Br J Cancer 90:283–288PubMedCrossRef

Avruch J, Khokhlatchev A, Kyriakis JM et al (2001) Ras activation of the Raf kinase: tyrosine kinase recruitment of the MAP kinase cascade. Recent Prog Horm Res 56:127–155PubMedCrossRef

Kolch W (2000) Meaningful relationships: the regulation of the Ras/Raf/MEK/ERK pathway by protein interactions. Biochem J 351(Pt 2):289–305PubMedCrossRef

Storm SM, Cleveland JL, Rapp UR (1990) Expression of raf family proto-oncogenes in normal mouse tissues. Oncogene 5:345–351PubMed

Barnier JV, Papin C, Eychène A, Lecoq O, Calothy G (1995) The mouse B-raf gene encodes multiple protein isoforms with tissue-specific expression. J Biol Chem 270:23381–23389PubMedCrossRef

Luckett JC, Hüser MB, Giagtzoglou N, Brown JE, Pritchard CA (2000) Expression of the A-raf proto-oncogene in the normal adult and embryonic mouse. Cell Growth Differ 11:163–171PubMed

Simister PC, Banfield MJ, Brady RL (2002) The crystal structure of PEBP-2, a homologue of the PEBP/RKIP family. Acta Crystallogr D Biol Crystallogr 58:1077–1080PubMedCrossRef

10.

Odabaei G, Chatterjee D, Jazirehi AR, Goodglick L, Yeung K, Bonavida B (2004) Raf-1 kinase inhibitor protein: structure, function, regulation of cell signaling, and pivotal role in apoptosis. Adv Cancer Res 91:169–200PubMedCrossRef

11.

Yeung K, Seitz T, Li S et al (1999) Suppression of Raf-1 kinase activity and MAP kinase signalling by RKIP. Nature 401:173–177PubMedCrossRef

12.

Park S, Yeung ML, Beach S, Shields JM, Yeung KC (2005) RKIP downregulates B-Raf kinase activity in melanoma cancer cells. Oncogene 24:3535–3540PubMedCrossRef

13.

Fu Z, Smith PC, Zhang L et al (2003) Effects of raf kinase inhibitor protein expression on suppression of prostate cancer metastasis. J Natl Cancer Inst 95:878–889PubMedCrossRef

14.

Schuierer MM, Bataille F, Hagan S, Kolch W, Bosserhoff AK (2004) Reduction in Raf kinase inhibitor protein expression is associated with increased Ras-extracellular signal-regulated kinase signaling in melanoma cell lines. Cancer Res 64:5186–5192PubMedCrossRef

15.

Van Gelder RN, von Zastrow ME, Yool A, Dement WC, Barchas JD, Eberwine JH (1990) Amplified synthesized from limited quantities of heterogeneous cDNA. Proc Natl Acad Sci USA 87:1663–1667PubMedCrossRef

16.

Diez Alvarez DR, Dopazo A (2007) Codelink: an R package for analysis of GE healthcare gene expression bioarrays. Bioinformatics 23:1168–1169CrossRef

17.

Altman NS, Hua J (2006) Extending the loop design for two-channel microarray experiments. Genet Res 88:153–163PubMedCrossRef

18.

Sturn A, Quackenbush J, Trajanoski Z (2002) Genesis: cluster analysis of microarray data. Bioinformatics 18:207–208PubMedCrossRef

19.

Boulalas I, Zaravinos A, Karyotis I, Delakas D, Spandidos DA (2009) Activation of RAS family genes in urothelial carcinoma. J Urol 181:2312–2319PubMedCrossRef

20.

Dyrskjøt L, Kruhøffer M, Thykjaer T et al (2004) Gene expression in the urinary bladder: a common carcinoma in situ gene expression signature exists disregarding histopathological classification. Cancer Res 64:4040–4048PubMedCrossRef

21.

Mengual L, Burset M, Ars E et al (2009) DNA microarray expression profiling of bladder cancer allows identification of noninvasive diagnostic markers. J Urol 182:741–748PubMedCrossRef

22.

Davies H, Bignell GR, Cox C et al (2002) Mutations of the BRAF gene in human cancer. Nature 417:949–954PubMedCrossRef

23.

Zlobec I, Kovac M, Erzberger P et al (2010) Combined analysis of specific KRAS mutation, BRAF and microsatellite instability identifies prognostic subgroups of sporadic and hereditary colorectal cancer. Int J Cancer Feb 16, 2010

24.

Zlobec I, Bihl MP, Schwarb H, Terracciano L, Lugli A (2009) Clinicopathological and protein characterization of BRAF- and K-RAS-mutated colorectal cancer and implications for prognosis. Int J Cancer Nov 11, 2009

25.

Libra M, Malaponte G, Navolanic PM et al (2005) Analysis of BRAF mutation in primary and metastatic melanoma. Cell Cycle 10:1382–1384CrossRef

26.

Baldus SE, Schaefer KL, Engers R, Hartleb D, Stoecklein NH, Gabbert HE (2010) Prevalence and heterogeneity of KRAS, BRAF, and PIK3CA mutations in primary colorectal adenocarcinomas and their corresponding metastases. Clin Cancer Res 16:790–799PubMedCrossRef

27.

Boulalas I, Zaravinos A, Delakas D, Spandidos DA (2009) Mutational analysis of the BRAF gene in transitional cell carcinoma of the bladder. Int J Biol Markers 24:17–21PubMed

28.

Zaravinos A, Kanellou P, Baritaki S, Bonavida B, Spandidos DA (2009) BRAF and RKIP are significantly decreased in cutaneous squamous cell carcinoma. Cell Cycle 8:1402–1408PubMedCrossRef

29.

Zaravinos A, Bizakis J, Spandidos DA (2008) RKIP and BRAF aberrations in human nasal polyps and the adjacent turbinate mucosae. Cancer Lett 264:288–298PubMedCrossRef

30.

Gattenlöhner S, Etschmann B, Riedmiller H, Müller-Hermelink HK (2009) Lack of KRAS and BRAF mutation in renal cell carcinoma. Eur Urol 55:1490–1491PubMedCrossRef

31.

Karlou M, Saetta AA, Korkolopoulou P et al (2009) Activation of extracellular regulated kinases (ERK1/2) predicts poor prognosis in urothelial bladder carcinoma and is not associated with B-Raf gene mutations. Pathology 41:327–334PubMedCrossRef

32.

Mhawech-Fauceglia P, Fischer G, Beck A, Cheney RT, Herrmann FR (2006) Raf1, Aurora-A/STK15 and E-cadherin biomarkers expression in patients with pTa/pT1 urothelial bladder carcinoma; a retrospective TMA study of 246 patients with long-term follow-up. Eur J Surg Oncol 32:439–444PubMedCrossRef

33.

McPhillips F, Mullen P, MacLeod KG et al (2006) Raf-1 is the predominant Raf isoform that mediates growth factor-stimulated growth in ovarian cancer cells. Carcinogenesis 27:729–739PubMedCrossRef

34.

Simon R, Richter J, Wagner U et al (2001) High-throughput tissue microarray analysis of 3p25 (RAF1) and 8p12 (FGFR1) copy number alterations in urinary bladder cancer. Cancer Res 61:4514–4519PubMed

35.

O’Neill E, Rushworth L, Baccarini M, Kolch W (2004) Role of the kinase MST2 in suppression of apoptosis by the proto-oncogene product Raf-1. Science 306:2267–2270PubMedCrossRef

36.

Ehrenreiter K, Piazzolla D, Velamoor V et al (2005) Raf-1 regulates Rho signaling and cell migration. J Cell Biol 168:955–964PubMedCrossRef

37.

Wan PT, Garnett MJ, Roe SM, Lee S et al (2004) Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF. Cell 116:855–867PubMedCrossRef

38.

Keller ET, Fu Z, Yeung K, Brennan M (2004) Raf kinase inhibitor protein: a prostate cancer metastasis suppressor gene. Cancer Lett 207:131–137PubMedCrossRef

39.

Fu Z, Kitagawa Y, Shen R et al (2006) Metastasis suppressor gene Raf kinase inhibitor protein (RKIP) is a novel prognostic marker in prostate cancer. Prostate 66:248–256PubMedCrossRef

40.

Blaveri E, Brewer JL, Roydasgupta R et al (2005) Bladder cancer stage and outcome by array-based comparative genomic hybridization. Clin Cancer Res 11:7012–7022PubMedCrossRef

41.

Keller ET, Fu Z, Brennan M (2005) The biology of a prostate cancer metastasis suppressor protein: Raf kinase inhibitor protein. J Cell Biochem 94:273–278PubMedCrossRef

42.

Woods Ignatoski KM, Grewal NK, Markwart SM et al (2008) Loss of Raf kinase inhibitory protein induces radioresistance in prostate cancer. Int J Radiat Oncol Biol Phys 72:153–160PubMedCrossRef

43.

Li HZ, Wang Y, Gao Y et al (2008) Effects of raf kinase inhibitor protein expression on metastasis and progression of human epithelial ovarian cancer. Mol Cancer Res 6:917–928PubMedCrossRef

44.

Lou TF, Gray CW, Gray DM (2003) The reduction of Raf-1 protein by phosphorothioate ODNs and siRNAs targeted to the same two mRNA sequences. Oligonucleotides 13:313–324PubMedCrossRef

45.

Chatterjee D, Bai Y, Wang Z et al (2004) RKIP sensitizes prostate and breast cancer cells to drug-induced apoptosis. J Biol Chem 279:17515–17523PubMedCrossRef

Title: Implication of RAF and RKIP Genes in Urinary Bladder Cancer
Authors: Apostolos Zaravinos
Maria Chatziioannou
George I. Lambrou
Ioannis Boulalas
Dimitris Delakas
Demetrios A. Spandidos
Publication date: 01-06-2011
Publisher: Springer Netherlands
Published in: Pathology & Oncology Research / Issue 2/2011
Print ISSN: 1219-4956
Electronic ISSN: 1532-2807
DOI: https://doi.org/10.1007/s12253-010-9295-1

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