Alternating expression levels of WWOX tumor suppressor and cancer-related genes in patients with bladder cancer

Płuciennik,Elżbieta; Nowakowska,Magdalena; Stępien,Anna; Wołkowicz,Mateusz; Stawiński,Adam; Różański,Waldemar; Lipiński,Marek; Bednarek,Andrzej  K

doi:10.3892/ol.2014.2476

Alternating expression levels of WWOX tumor suppressor and cancer-related genes in patients with bladder cancer

Authors:
- Elżbieta Płuciennik
- Magdalena Nowakowska
- Anna Stępien
- Mateusz Wołkowicz
- Adam Stawiński
- Waldemar Różański
- Marek Lipiński
- Andrzej K Bednarek
View Affiliations

Affiliations: Department of Molecular Cancerogenesis, Medical University of Lodz, Lodz 90-752, Poland, Laboratory of Clinical and Transplant Immunology and Genetics, Copernicus Memorial Hospital in Lodz, Lodz 93‑513, Poland, Bio-Tech Consulting Ltd., Lodz 90‑212; 4Second Department of Urology, Medical University of Lodz, Lodz 93‑513, Poland, Second Department of Urology, Medical University of Lodz, Copernicus Memorial Hospital in Lodz, Lodz 93‑513, Poland
Published online on: August 22, 2014 https://doi.org/10.3892/ol.2014.2476
Pages: 2291-2297

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to determine the roles of the WWOX tumor suppressor and cancer-related genes in bladder tumor carcinogenesis. Reverse transcription‑quantitative polymerase chain reaction was used to analyze the status of WWOX promoter methylation (using MethylScreen™ technology) and loss of heterozygosity (LOH) in papillary urothelial cancer tissues. The associations between the expression levels of the following tumorigenesis‑related genes were also assessed: The WWOX tumor suppressor gene, the MKI67 proliferation gene, the BAX, BCL2 and BIRC5 apoptotic genes, the EGFR signal transduction gene, the VEGF vascular endothelial growth factor gene, and the CCND1 and CCNE1 cell cycle genes. The results reveal a high frequency of LOH in intron 1 in the WWOX gene, as well as an association between reduced WWOX expression levels and increased promoter methylation. In addition, the present study demonstrates that in bladder tumors, apoptosis is inhibited by increased expression levels of the BCL2 gene. A correlation between the proliferation indices of the MKI67 and the BIRC5 genes was also revealed. Furthermore, the expression levels of VEGF were identified to be positively associated with those of the EGFR gene.

View Figures

View References

1	Ministry of Health, Poland. National Cancer Registry. http://epid.coi.waw.pl/krn/. Accessed June 30, 2010
2	Luis NM, López-Knowles E and Real FX: Molecular biology of bladder cancer. Clin Transl Oncol. 9:5–12. 2007.
3	Kim WJ and Bae SC: Molecular biomarkers in urothelial bladder cancer. Cancer Sci. 99:646–652. 2008.
4	Jung I and Messing E: Molecular mechanisms and pathways in bladder cancer development and progression. Cancer Control. 7:325–334. 2000.
5	Mitra AP, Datar RH and Cote RJ: Molecular pathways in invasive bladder cancer: new insights into mechanisms, progression, and target identification. J Clin Oncol. 24:5552–5564. 2006.
6	Yoon DS, Li L, Zhang RD, Kram A, et al: Genetic mapping and DNA sequence-based analysis of deleted regions on chromosome 16 involved in progression of bladder cancer from occult preneoplastic conditions to invasive disease. Oncogene. 20:5005–5014. 2001.
7	Bednarek AK, Laflin KJ, Daniel RL, et al: WWOX, a novel WW domain-containing protein mapping to human chromosome 16q23.3–24.1, a region frequently affected in breast cancer. Cancer Res. 60:2140–2145. 2000.
8	Qin HR, Iliopoulos D, Semba S, et al: A role for the WWOX gene in prostate cancer. Cancer Res. 66:6477–6481. 2006.
9	Lan C, Chenggang W, Yulan B, et al: Aberrant expression of WWOX protein in epithelial ovarian cancer: a clinicopathologic and immunohistochemical study. Int J Gynecol Pathol. 31:125–132. 2012.
10	Yang W, Cui S, Ma J, et al: Cigarette smoking extract causes hypermethylation and inactivation of WWOX gene in T-24 human bladder cancer cells. Neoplasma. 59:216–223. 2012.
11	Ramos D, Abba M, Lopez-Guerrero JA, et al: Low levels of WWOX protein immunoexpression correlate with tumour grade and a less favourable outcome in patients with urinary bladder tumours. Histopathology. 52:831–839. 2008.
12	Iliopoulos D, Guler G, Han SY, et al: Fragile genes as biomarkers: epigenetic control of WWOX and FHIT in lung, breast and bladder cancer. Oncogene. 24:1625–1633. 2005.
13	Knudson AG Jr: Hereditary cancer, oncogenes, and antioncogenes. Cancer Res. 45:1437–1443. 1985.
14	Kuroki T, Yendamuri S, Trapasso F, et al: The tumor suppressor gene WWOX at FRA16D is involved in pancreatic carcinogenesis. Clin Cancer Res. 10:2459–2465. 2004.
15	Kuroki T, Trapasso F, Shiraishi T, et al: Genetic alterations of the tumor suppressor gene WWOX in esophageal squamous cell carcinoma. Cancer Res. 62:2258–2260. 2002.
16	Aqeilan RI, Donati V, Palamarchuk A, et al: WW domain-containing proteins, WWOX and YAP, compete for interaction with ErbB-4 and modulate its transcriptional function. Cancer Res. 65:6764–6772. 2005.
17	Aqeilan RI, Donati V, Gaudio E, et al: Association of Wwox with ErbB4 in breast cancer. Cancer Res. 67:9330–9336. 2007.
18	Gaudio E, Palamarchuk A, Palumbo T, et al: Physical association with WWOX suppresses c-Jun transcriptional activity. Cancer Res. 66:11585–11589. 2006.
19	Aqeilan RI, Pekarsky Y, Herrero JJ, et al: Functional association between Wwox tumor suppressor protein and p73, a p53 homolog. Proc Natl Acad Sci USA. 101:4401–4406. 2004.
20	Aqeilan RI, Hassan MQ, de Bruin A, et al: The WWOX tumor suppressor is essential for postnatal survival and normal bone metabolism. J Biol Chem. 283:21629–21639. 2008.
21	Jin C, Ge L, Ding X, et al: PKA-mediated protein phosphorylation regulates ezrin-WWOX interaction. Biochem Biophys Res Commun. 341:784–791. 2006.
22	Nunez MI, Ludes-Meyers J and Aldaz CM: WWOX protein expression in normal human tissues. J Mol Histol. 37:115–125. 2006.
23	Aqeilan RI, Hagan JP, de Bruin A, et al: Targeted ablation of the WW domain-containing oxidoreductase tumor suppressor leads to impaired steroidogenesis. Endocrinology. 150:1530–1535. 2009.
24	World Health Organization Classification of Tumours. World Health Organization, IARC Press; Lyon: 2004
25	Kosla K, Pluciennik E, Kurzyk A, Jesionek-Kupnicka D, et al: Molecular analysis of WWOX expression correlation with proliferation and apoptosis in glioblastoma multiforme. J Neurooncol. 101:207–213. 2011.
26	Płuciennik E, Nowakowska M, Wujcicka WI, et al: Genetic alterations of WWOX in Wilms’ tumor are involved in its carcinogenesis. Oncol Rep. 28:1417–1422. 2012.
27	Pfaffl MW, Horgan GW and Dempfle L: Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res. 30:e362002.
28	Maeda N, Semba S, Nakayama S, et al: Loss of WW domain-containing oxidoreductase expression in the progression and development of gastric carcinoma: clinical and histopathologic correlations. Virchows Arch. 457:423–432. 2010.
29	Aqeilan RI, Kuroki T, Pekarsky Y, et al: Loss of WWOX expression in gastric carcinoma. Clin Cancer Res. 10:3053–3058. 2004.
30	Yendamuri S, Kuroki T, Trapasso F, et al: WW domain containing oxidoreductase gene expression is altered in non-small cell lung cancer. Cancer Res. 63:878–881. 2003.
31	Chen T, Sahin A and Aldaz CM: Deletion map of chromosome 16q in ductal carcinoma in situ of the breast: refining a putative tumor suppressor gene region. Cancer Res. 56:5605–5609. 1996.
32	Shariat SF, Ashfaq R, Sagalowsky AI and Lotan Y: Correlation of cyclin D1 and E1 expression with bladder cancer presence, invasion, progression, and metastasis. Hum Pathol. 37:1568–1576. 2006.
33	Shiina H, Igawa M, Urakami S, et al: Immunohistochemical analysis of Bcl-2 expression in transitional cell carcinoma of the bladder. J Clin Pathol. 49:395–399. 1996.
34	Bilim VN, Tomita Y, Kawasaki T, et al: Variable Bcl-2 phenotype in benign and malignant lesions of urothelium. Cancer Lett. 128:87–92. 1998.
35	Wang H, Xi X, Kong X, et al: The expression and significance of survivin mRNA in urinary bladder carcinomas. J Cancer Res Clin Oncol. 130:487–490. 2004.
36	Shariat SF, Ashfaq R, Karakiewicz PI, et al: Survivin expression is associated with bladder cancer presence, stage, progression, and mortality. Cancer. 109:1106–1113. 2007.
37	Kitamur H, Torigoe T, Honma I, et al: Expression and antigenicity of survivin, an inhibitor of apoptosis family member, in bladder cancer: Implications for specific immunotherapy. Urology. 67:955–959. 2006.
38	Gonzalez-Campora R, Davalos-Casanova G, Beato-Moreno A, et al: Apoptotic and proliferation indexes in primary superficial bladder tumors. Cancer Lett. 242:266–272. 2006.
39	Yang CC, Chu KC and Yeh WM: The expression of vascular endothelial growth factor in transitional cell carcinoma of urinary bladder is correlated with cancer progression. Urol Oncol. 22:1–6. 2004.
40	Crew JP, O’Brien T, Bradburn M, et al: Vascular endothelial growth factor is a predictor of relapse and stage progression in superficial bladder cancer. Cancer Res. 57:5281–5285. 1997.
41	Pfister C, Moore L, Allard P, Larue H, Lacombe L, Têtu B, Meyer F and Fradet Y: Predictive value of cell cycle markers p53, MDM2, p21, and Ki-67 in superficial bladder tumor recurrence. Clin Cancer Res. 5:4079–4084. 1999.
42	Behnsawy HM, Miyake H, Abdalla MA, Sayed MA, Ahmed Ael-F and Fujisawa M: Expression of cell cycle-associated proteins in non-muscle-invasive bladder cancer: correlation with intravesical recurrence following transurethral resection. Urol Oncol. 29:495–501. 2011.
43	Chen L, Wang X, Mei H and Chen W: Apoptosis and expression of PCNA in superficial transitional cell bladder cancer as related to recurrence. Zhonghua Wai Ke Za Zhi. 36:484–486. 1998.(In Chinese).
44	Gazzaniga P, Gradilone A, Giuliani L, Gandini O, Silvestri I, Nofroni I, Saccani G, Frati L and Aglianò AM: Expression and prognostic significance of LIVIN, SURVIVIN and other apoptosis-related genes in the progression of superficial bladder cancer. Ann Oncol. 14:85–90. 2003.
45	Jeong IG, Kim SH, Jeon HG, Kim BH, Moon KC, Lee SE and Lee E: Prognostic value of apoptosis-related markers in urothelial cancer of the upper urinary tract. Hum Pathol. 40:668–677. 2009.
46	Chow NH, Liu HS, Lee EI, Chang CJ, Chan SH, Cheng HL, Tzai TS and Lin JS: Significance of urinary epidermal growth factor and its receptor expression in human bladder cancer. Anticancer Res. 17:1293–1296. 1997.
47	Neal DE, Marsh C, Bennett MK, et al: Epidermal-growth-factor receptors in human bladder cancer: comparison of invasive and superficial tumours. Lancet. 1:366–368. 1985.