Korean J Urol.  2007 Apr;48(4):402-407. 10.4111/kju.2007.48.4.402.

Clusterin Expression and Apoptosis in Transitional Cell Carcinoma of the Bladder

• Park HJ
• Lee HJ
• Yum YH
• Kang JY
• Yoo TK

• KMID: 1997107
• DOI: http://doi.org/10.4111/kju.2007.48.4.402

Abstract

PURPOSE
The clusterin expression has been associated with tumorigenesis of various malignancies, including tumors of the prostate, colon and breast. Furthermore, the expression of clusterin is modulated by many factors that are believed to regulate tumor growth and apoptosis. We studied the clusterin expression in transitional cell carcinoma (TCC) of the bladder and we investigated its correlation with apoptosis.
MATERIALS AND METHODS
Eighty five bladder tumor specimens from radical cystectomy or transurethral resection were subjected to immunohistochemical clusterin staining with Ig G clusterin Ab. We examined the immunohistochemical localization of clusterin, and this was followed by TUNEL staining to detect the apoptotic cells. After double-staining with Hoechst 33258, we detected the apoptotic cells under a fluorescence microscope and we calculated the apoptotic index.
RESULTS
Invasive TCC showed a stronger positive expression of clusterin as compared with superficial TCC, but the positivity of the clusterin expression was not in proportion to the tumor grade. The apoptotic indices of cancer were 0.52+/-0.28%, 0.30+/-0.16% and 0.17+/-0.11% in Grade I, Grade II and Grade III superficial TCC, respectively, and it was 0.23+/-0.13% in Grade III invasive TCC. Apoptotic cells were not detected in the cancer cells stained with clusterin. Conversely, clusterin was not expressed in the cells showing apoptosis.
CONCLUSIONS
These results suggest that clusterin could be used as a marker to provide prognostic information for the TCC patients. The apoptotic index revealed that apoptosis and the clusterin expression have correlation with transitional cell cancer. Further study will be needed to clarify the role of clusterin as a therapeutic target for cancer treatment.

Keyword

Apoptosis; Clusterin; Transitional cell carcinoma

MeSH Terms

Apoptosis*
Bisbenzimidazole
Breast
Carcinogenesis
Carcinoma, Transitional Cell*
Clusterin*
Colon
Cystectomy
Fluorescence
Humans
In Situ Nick-End Labeling
Prostate
Urinary Bladder Neoplasms
Urinary Bladder*
Bisbenzimidazole
Clusterin

Figure

• 

  Fig. 1 The expressions of clusterin in bladder cancer specimens with the degree, from negative (A) to 3+ (D).

• 

  Fig. 2 The clusterin expression is shown (A) and the same slide stained with TUNEL assay (B). Cells stained with clusterin are not stained with TUNEL (C).

Reference

1. Trougakos IP, Gonos ES. Clusterin/apolipoprotein J in human aging and cancer. Int J Biochem Cell Biol. 2002. 34:1430–1448.

2. Trougakos IP, So A, Jansen B, Gleave ME, Gonos ES. Silencing expression of the clusterin/apolipoprotein j gene in human cancer cells using small interfering RNA induces spontaneous apoptosis, reduced growth ability, and cell sensitization to genotoxic and oxidative stress. Cancer Res. 2004. 64:1834–1842.

3. Cheng CY, Chen CL, Feng ZM, Marshall A, Bardin CW. Rat clusterin isolated from primary Sertoli cell-enriched culture medium is sulfated glycoprotein-2 (SGP-2). Biochem Biophys Res Commun. 1988. 155:398–404.

4. Fritz IB, Burdzy K, Setchell B, Blaschuk O. Ram rete testis fluid contains a protein (clusterin) which influences cell-cell interactions in vitro. Biol Reprod. 1983. 28:1173–1188.

5. Sensibar JA, Sutkowski DM, Raffo A, Buttyan R, Griswold MD, Sylvester SR, et al. Prevention of cell death induced by tumor necrosis factor alpha in LNCaP cells by overexpression of sulfated glycoprotein-2 (clusterin). Cancer Res. 1995. 55:2431–2437.

6. Flanagan L, Van Weelden K, Ammerman C, Ethier SP, Welsh J. SUM-159PT cells: a novel estrogen independent human breast cancer model system. Breast Cancer Res Treat. 1999. 58:193–204.

7. Leskov KS, Klokov DY, Li J, Kinsella TJ, Boothman DA. Synthesis and functional analyses of nuclear clusterin, a cell death protein. J Biol Chem. 2003. 278:11590–11600.

8. Miyake H, Gleave M, Kamidono S, Hara I. Overexpression of clusterin in transitional cell carcinoma of the bladder is related to disease progression and recurrence. Urology. 2002. 59:150–154.

9. Miyake H, Hara I, Kamidono S, Gleave ME. Synergistic chemsensitization and inhibition of tumor growth and metastasis by the antisense oligodeoxynucleotide targeting clusterin gene in a human bladder cancer model. Clin Cancer Res. 2001. 7:4245–4252.

10. Kruger S, Mahnken A, Kausch I, Feller AC. Value of clusterin immunoreactivity as a predictive factor in muscle-invasive urothelial bladder carcinoma. Urology. 2006. 67:105–109.

11. July LV, Beraldi E, So A, Fazli L, Evans K, English JC, et al. Nucleotide-based therapies targeting clusterin chemosensitize human lung adenocarcinoma cells both in vitro and in vivo. Mol Cancer Ther. 2004. 3:223–232.

12. Chen X, Halberg RB, Ehrhardt WM, Torrealba J, Dove WF. Clusterin as a biomarker in murine and human intestinal neoplasia. Proc Natl Acad Sci USA. 2003. 100:9530–9535.

13. Xie MJ, Motoo Y, Su SB, Mouri H, Ohtsubo K, Matsubara F, et al. Expression of clusterin in human pancreatic cancer. Pancreas. 2002. 25:234–238.

14. Zellweger T, Kiyama S, Chi K, Miyake H, Adomat H, Skov K, et al. Overexpression of the cytoprotective protein clusterin decreases radiosensitivity in the human LNCaP prostate tumour model. BJU Int. 2003. 92:463–469.

15. Miyake H, Hara I, Kamidono S, Gleave ME, Eto H. Resistance to cytotoxic chemotherapy-induced apoptosis in human prostate cancer cells is associated with intracellular clusterin expression. Oncol Rep. 2003. 10:469–473.

16. Scaltriti M, Brausi M, Amorosi A, Caporali A, D'Arca D, Astancolle S, et al. Clusterin (SGP-2, ApoJ) expression is downregulated in low- and high-grade human prostate cancer. Int J Cancer. 2004. 108:23–30.

17. Pucci S, Bonanno E, Pichiorri F, Angeloni C, Spagnoli LG. Modulation of different clusterin isoforms in human colon tumorigenesis. Oncogene. 2004. 23:2298–2304.

18. Yoon JH, Lee JH, Yeom BW, Won NH, Yoon DK. Expression of osteopontin and clusterin in transitional cell carcinoma of the bladder. Korean J Urol. 2005. 46:341–346.

19. Steinberg J, Oyasu R, Lang S, Sintich S, Rademaker A, Lee C, et al. Intracellular levels of SGP-2 (Clusterin) correlate with tumor grade in prostate cancer. Clin Cancer Res. 1997. 3:1707–1711.

20. Redondo M, Villar E, Torres-Munoz J, Tellez T, Morell M, Petito CK. Overexpression of clusterin in human breast carcinoma. Am J Pathol. 2000. 157:393–399.

21. Miyake H, Hara S, Zellweger T, Kamidono S, Gleave ME, Hara I. Acquisition of resistance to Fas-mediated apoptosis by overexpression of clusterin in human renal-cell carcinoma cells. Mol Urol. 2001. 5:105–111.

22. Criswell T, Klokov D, Beman M, Lavik JP, Boothman DA. Repression of IR-inducible clusterin expression by the p53 tumor suppressor protein. Cancer Biol Ther. 2003. 2:372–380.