|Year : 2009 | Volume
| Issue : 2 | Page : 155-158
|Immunohistochemical positive stained p53 protein in bladder transitional cell carcinoma
Monireh Halimi1, Alireza Salehi1, Hosein Baybordi1, Nariman Nezami2
1 Department of Pathology, Tabriz University of Medical Sciences, Tabriz, Iran
2 Young Researchers Club, Tabriz Islamic Azad University, Tabriz; Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Click here for correspondence address and email
| Abstract|| |
Background: Molecular genetics and immunopathologic analysis of bladder cancer have shown some abnormalities in a number of genes and proteins that have been implicated in the development and progression of such tumors, mainly in the p53 pathway. Aims: To investigate the rate of positively stained p53 protein in patients with urothelial papillary carcinoma of the bladder (UCB) by immunohistochemistry and its relationship with tumor grade, gender and age of the patients. Settings and Design: During the present cross-sectional study, 100 paraffin-embedded specimens of UCB, which were provided from biopsies of the bladder by transurethral access, were immunohistochemically stained and studied for p53 protein from May 2006 to May 2007 in our referral center pathology laboratory. Materials and Methods: First, 4 µm slices of paraffin sections were provided and then stained by the avidin-biotin peroxidase method. The rate of positively stained p53 protein (defined as positive nuclear staining in over 10% of the cells) was assessed. This rate was also estimated and compared between grades, genders and age-related groups (<70 years, ≥70 years). Statistical Analysis: The χ2 , Fisher's exact test and Mann-Whitney U test were used for comparing. Results: The overall rate of positively stained specimens was 11% for nuclear p53 protein. This rate was significantly higher in females (10/29 vs. 1/71; P < 0.001; odds ratio [OR]: 0.23; 95% confidence interval [CI]: 4.43-306.08), patients with 70 or older than 70 years (8/42 vs. 3/58; P = 0.04; OR: 0.55; 95% CI: 1.07-17.39) and in high-grade tumors (10/58 vs. 1/42; P = 0.02; OR: 0.59; 95% CI: 0.01-0.95). Conclusions: The rate of positively stained p53 protein for UCB was lower in our population. This rate was also higher in females, patients with 70 or older than 70 years and high grade of UCB.
Keywords: Bladder, immunohistochemistry, p53, transitional cell carcinoma
|How to cite this article:|
Halimi M, Salehi A, Baybordi H, Nezami N. Immunohistochemical positive stained p53 protein in bladder transitional cell carcinoma. Indian J Pathol Microbiol 2009;52:155-8
|How to cite this URL:|
Halimi M, Salehi A, Baybordi H, Nezami N. Immunohistochemical positive stained p53 protein in bladder transitional cell carcinoma. Indian J Pathol Microbiol [serial online] 2009 [cited 2019 Sep 20];52:155-8. Available from: http://www.ijpmonline.org/text.asp?2009/52/2/155/48905
| Introduction|| |
Bladder cancer is one of the most common malignancies occurring worldwide.  These tumors are believed to arise as a consequence of irreversible damages of the DNA (initiation), continued division and proliferation (promotion). Progression toward neoplasia may require cumulative effects of one or more initiating/promoting agents.  Molecular genetics and immunopathologic analysis of bladder cancer have identified a number of abnormalities in some of the genes and proteins that have been implicated in the development and progression of such tumors, mainly in the p53 pathway. ,,
The tumor suppressor gene p53 is located on chromosome 17p13 and encodes a nuclear phosphoprotein involved in the cell cycle that allows cellular DNA repair and/or apoptosis, which occur by controlling cellular progression from the G1 to the S phase. ,
Mutant p53 has a prolonged half-life and can be demonstrated by immunohistochemical techniques.  Overexpression of p53 has been correlated with the grade of bladder cancer. ,,,,
This study aimed to evaluate the rate of positive immunohistochemically stained protein of p53 in samples of urothelial papillary carcinoma of the bladder (UCB) and its relationship with the grade of malignancy, gender and age of the patients.
| Materials and Methods|| |
During this retrospective cross-sectional study, 100 paraffin-embedded blocks of UCB specimens were randomly selected and immunohistochemically stained for overexpression of p53 (by a kit from Dako Cytomation California Inc., Carpentaria, USA; Lot no: 10468) during a 1-year period from May 2006 to May 2007 in the pathology laboratory of our referral hospital. These UCB specimens were provided by transurethral access and archived in the pathology laboratory over a 1-year period. Four micron slices of paraffin sections were provided and stained by the avidin-biotin peroxidase method. A light microscope (Nikon, Tokyo, Japan) was used for the evaluation of the stained specimens.
Staining was considered positive when over 10% of the cell nuclei had been stained. , The percentages of tumor cells with positive p53 protein staining were determined independently by two researchers (M.H., H.B.) in 300-1000 tumor cells. Quotients (positive tumor cells/total counted tumor cells) were calculated as percentage and rounded to the nearest integer. The arithmetic mean of both observers' scores was used for statistical analysis.  The grade of tumors (low or high grade) was also determined using the WHO/ISUP system.  The rate of positivity was compared between the genders, grades of UCB (low or high) and the patients younger than 70 years and 70 years old or older. The mean age of the patients was compared between the participants with positive and negative staining results for the p53 protein.
Data were analyzed using the SPSS statistical software package (version 11.0; SPSS Inc., Chicago, IL, USA). Continuous variables were demonstrated as mean ± standard deviation (SD) and categorical data were shown as frequency and percent. The χ2 , Fisher's exact test and Mann-Whitney U test were applied for comparisons. P -values < 0.05 were considered statistically significant. The odds ratio (OR) and the 95% confidence interval (CI) were used to estimate the risk.
| Results|| |
Finally, paraffin-embedded sections of 100 participants with UCB were analyzed. Seventy-one and 29% of the patients were male and female, respectively. The mean age of the participants was 64.69 ± 12.99 years (35-84 years). Fifty-eight (58%) of participants were younger than 70 years and the remaining 42 (42%) were 70 or older than 70 years. Forty-two (42%) and 58 (58%) of the specimens were high-grade and low-grade UCB, respectively.
Eleven percent of the specimens were positively stained for nuclear p53 protein and 89% were not. [Figure 1a] and [Figure 1b] demonstrate the p53 protein positive staining in low- (10×) and high- (40×) power views, respectively. [Figure 2] demonstrates the hematoxylin and eosin staining of UCB low and high grades in low- (×100) and high- (×400) power views, respectively. The frequency of p53 protein positive staining results according to the age decades has been shown in [Figure 3].
As shown in [Table 1], the rates of positive staining for nuclear p53 protein were significantly higher in females ( P < 0.001; OR: 0.23; 95% CI: 4.43-306.08), patients with 70 or older than 70 years ( P = 0.04; OR: 0.55; 95% CI: 1.07-17.39) and in high grade of tumors ( P = 0.02; OR: 0.59; 95% CI: 0.01-0.95). The mean age of the patients with a positive staining result was significantly higher than the negatively stained results (69.81 ± 17.11 vs. 64.05 ± 12.36 years; P = 0.04, z = -2.01).
| Discussion|| |
Bladder neoplasms show a wide variation in the natural course and prognosis. Determination of specific biochemical markers for the changes resulting from the process of carcinogenesis offers an attractive approach to identify individuals at a risk of bladder cancer. 
During the last few years, mutation of the tumor suppressor gene p53 has been most widely studied as a genetic defect in bladder cancer.  Beside the many diverse functions of p53 protein, it acts as a factor that upregulates the activity of several genes modulating apoptosis.  However, the mutant p53 inhibits apoptosis. 
There is a diverse range of reports for positive staining of p53 protein in patients with TCC, from 7 to 78%. ,,,,,,,,,,,,,, In the present study, 11% of the specimens were positively stained for p53 protein. This rate is in the reported range, but it is closer to the lower limit. Variation in results may be due to the different assay methods, including the choice of anti-p53 antibody, immunohistochemistry protocol and scoring criteria.  Long standing of specimens in paraffin may have also contributed to the lower rate of positive staining in our study.
During the study of Al-Abadi et al. on 147 patients with TCC, frozen sections of tumor biopsies were obtained by transurethral resection and then immunohistochemically stained using the monoclonal antibody clone D0-7 (DAKO).  Like in our study, tumors expressing p53 in more than 10% of the nuclei were regarded positive, which was found in 84 (57%) of the 147 examined tumors.
Despite the methodological difference, in consistency with the Lin et al.  study, the rates of positive staining for nuclear p53 protein in the present study were higher in females. Lin et al. have reported p53 gene mutation in 33/54 males and 14/21 females, although the correlation was not studied.
Positive p53 staining was reported in 10-75% of low-grade tumors and in more than 58% of high-grade tumors. ,,,,, In a study by Ye et al. , positive nuclear staining for p53 was shown in 34 (51%) of 67 patients with TCC.  Also, Ye et al. have reported 40.8% and 78% p53 protein positive staining in low- and high-grade tumors, respectively ( P < 0.01).  Moch et al. showed a similar difference between the low- and high-grade tumors of UCB ( P < 0.0001).  Most of the previous studies have demonstrated that the rate of p53 expression in the patients with high-grade tumor is higher in than patients with low-grade tumor. ,,,, The present study results were in consistency with these studies, i.e. a higher rate of p53 protein-positive staining was seen in the high-grade tumor.
p53 protein is considered as a prognostic marker that might reflect the potent of aggressive malignancy, and poor prognosis,  and the results of the current study confirmed it by demonstrating a higher rate of p53 protein-positive staining in high-grade UCB.
In conclusion, the rate of positively stained p53 protein in UCB was lower in the population of our region (northwest of Iran). This rate was also higher for females, patients with 70 or older than 70 years and high-grade UCB.
| References|| |
|1.||Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, et al . Cancer statistics, 2008. CA Cancer J Clin 2008;58:71-96. |
|2.||Shankey TV. Urological cancer. In: Bauer KD, Duque RE, Shankey TV, editors. Clinical flow cytometry: Principles and application. 1 st ed. Philadelphia: Williams and Wilkins; 1993. p. 117-42. |
|3.||Ryan KM, Phillips AC, Vousden KH. Regulation and function of the p53 tumor suppressor protein. Curr Opin Cell Biol 2001;13:332-7. |
|4.||Reznikoff CA, Belair CD, Yeager TR, Savelieva E, Blelloch RH, Puthenveettil JA, et al . A molecular genetics model of human bladder cancer pathogenesis. Semin Oncol 1996;23:571-84. |
|5.||Cote RJ, Chatterjee SJ. Molecular determinants of outcome in bladder cancer. Cancer J Sci Am 1999;5:2-15. |
|6.||Greenblatt MS, Bennett WP, Hollstein M, Harris CC. Mutations with p53 tumor suppressor gene: Clues to cancer etiology and molecular pathogenesis. Cancer Res 1994;54:4855-78. |
|7.||Harris CC. Structure and function of the p53 tumor suppressor gene: Clues for rational cancer therapeutic strategies. J Natl Cancer Inst 1996;88:1442-55. |
|8.||Hall PA, Ray A, Lemione NR, Midgely CA, Krausz I, Lan DP. P53 immunostaining as a marker of malignant disease in diagnostic cytopathology. Lancet 1991;338:513. |
|9.||Sarkis AS, Bajorin DF, Reuter VE, Herr HV, Netto G, Zhang ZF, et al . Prognostic value of p53 nuclear over expression in patients with invasive bladder cancer treated with neoadjuvant MVAC. J Clin Oncol 1995;13:1384-90. |
|10.||Wright C, Mellon K, Johnstone P, Lane DP, Harris AL, Horne CH, et al . Expression of mutant p53, c-erbB-2 and the epidermal growth factor receptor in transitional cell carcinoma of the human urinary bladder. Br J Cancer 1991;63:967-70. |
|11.||Murphy WM. Current status of urinary cytology in the evaluation of bladder neoplasms. Hum Pathol 1990;21:886-96. |
|12.||Fujimoto K, Yamada Y, Okajima E, Kakizoe T, Sasaki H, Sugimura T, et al . Frequent association of p53 gene mutation in invasive bladder cancer. Cancer Res 1992;52:1393-8. |
|13.||Cesarman E, Chadburn A, Inghirami G, Gaidano G, Knowles DM. High levels of p53 protein expression do not correlate with p53 gene mutation in anaplastic large cell lymphoma. Am J Pathol 1993;143:845-56. |
|14.||Chatterjee SJ, Datar R, Youssefzadeh D, George B, Goebell PJ, Stein JP, et al . Combined effects of p53, p21, and pRb expression in the progression of bladder transitional cell carcinoma. J Clin Oncol 2004;22:975-7. |
|15.||Zigeuner R, Tsybrovskyy O, Ratschek M, Rehak P, Lipsky K, Langner C. Prognostic impact of p63 and p53 expression in upper urinary tract transitional cell carcinoma. Urology 2004;63:1079-83. |
|16.||Fernαndez-Figueras MT, Puig L, Musulen E, Gilaberte M, Ferrαndiz C, Lerma E, Ariza A. Prognostic significance of p27Kip1, p45Skp2 and Ki67 expression profiles in Merkel cell carcinoma, extracutaneous small cell carcinoma and cutaneous squamous cell carcinoma. Histopathology 2005;46:614-21. |
|17.||Ordonez NG, Rosai J. Urinary tract: Kidney, renal pelvis, and ureter bladder. In: Rosai J, editor. Rosai and Ackerman's surgical pathology. 9 th ed. Philadelphia: Elsevier; 2004. p. 1333. |
|18.||Bane BL, Rao JY, Hernstreet GP. Pathology and staging of bladder cancer. Semin Oncol 1996;23:546-70. |
|19.||Esring D, Elmaijan D, Groshen S, Freeman JA, Stein JP, Chen S, et al . Accumulation of nuclear p53 and tumor progression in bladder cancer. N Engl J Med 1994;331:1259-64. |
|20.||Bargonetti J, Manfredi JJ. Multiple roles of the tumor suppressor p53. Curr Opin Oncol 2002;14:86-91. |
|21.||Lu QL, Abel P, Foster CS, Lalani EN. Bcl-2: Role in epithelial differentiation and oncogenesis. Hum Pathol 1996;27:102-10. |
|22.||Burkhard FC, Markwalder R, Thalmann GN, Studer UE. Immunohistochemical determination of p53 over expression: An easy and readily available method to identify progression in superficial bladder cancer? Urol Res 1997;25:S31-5. |
|23.||Ye DW, Zheng JF, Qian SX, Ma YJ. Expression of p53 product in Chinese human bladder carcinoma. Urol Res 1993;21:223-6. |
|24.||Grossman HB, Liebert M, Antelo M, Dinney CP, Hu SX, Palmer JL, et al . p53 and RB expression predict progression in T1 bladder cancer. Clin Cancer Res 1998;4:829-34. |
|25.||Leonard C, Huret JL, Gfco; Groupe franηais de cytogιnιtique oncologique. From cytogenetics to cytogenomics of bladder cancers. Bull Cancer 2002;89:166-73. |
|26.||Gil P, Allepuz C, Blas M, Borque A, Del-Agua C, Plaza L, et al . Significance of protein p53 over expression in the clinical course of high-risk superficial bladder cancer. Urol Int 2003;70:172-7. |
|27.||Toktas G, Turkeri LN, Unluer E, Caliskan M, Aksoy B, Akdas A. Clinical significance of nuclear p53 protein accumulation in bladder cancer. Int Urol Nephrol 1999;31:327-34. |
|28.||Nakopoulou L, Constantinides C, Papandropoulos J, Theodoropoulos G, Tzonou A, Giannopoulos A, et al . Evaluation of over expression of p53 tumor suppressor protein in superficial and invasive transitional cell bladder cancer, comparison with DNA ploidy. Urology 1995;46:334-40. |
|29.||Nakopoulou L, Vourlakou C, Zervas A, Tzonou A, Gakiopoulou H, Dimopoulos MA. The prevalence of bcl-2, p53, and Ki-67 immunoreactivity in transitional cell bladder carcinomas and their clinicopathologic correlates. Hum Pathol 1998;29:146-54. |
|30.||Uchida T, Minei S, Gao JP, Wang C, Satoh T, Baba S. Clinical significance of p53, MDM2 and bcl-2 expression in transitional cell carcinoma of the bladder. Oncol Rep 2002;9:253-9. |
|31.||Babjuk M, Soukup V, Mares J, Duskova J, Sedlacek Z, Trkova M, et al . The expression of PAX5, p53 immunohistochemistry and p53 mutation analysis in superficial bladder carcinoma tissue: Correlation with pathological findings and clinical outcome. Int Urol Nephrol 2002-2003;34:495-501. |
|32.||Rebassa Llull M, Gutiιrrez Sanz-Gadea C, Muρoz Vιlez D, Esteva Bauzα M, Gómez Bellvert C, Mus Malleu A, et al . Expression of protein p53 in superficial transitional carcinoma of the bladder with differing course. Arch Esp Urol 1995;52:1061-4. |
|33.||Inagaki T, Ebisuno S, Uekado Y, Hirano A, Hiroi A, Shinka T, et al . PCNA and p53 in urinary bladder cancer: Correlation with histological findings and prognosis. Int J Urol 1997;4:172-7. |
|34.||Thomas DJ, Robinson MC, Charlton R, Wilkinson S, Shenton BK, Neal DE. p53 expression, ploidy and progression in pT1 transitional cell carcinoma of the bladder. Br J Urol 1994;73:533-7. |
|35.||Abbal-Helmy WH, Selmy G. Evaluation of P53, bcl-2 proteins and DNA ploidy in squamous and transitional cell carcinomas of urinary bladder. J Egypt Nat Cancer Inst 2001;12:293-9. |
|36.||Amirghofran Z, Monabati A, Khezri A, Malek-Hosseini Z. Apoptosis in transitional cell carcinoma of bladder and its relation to proliferation and expression of p53 and Bcl-2. Pathol Oncol Res 2004;10:154-8. |
|37.||Shi SR, Cote RJ, Taylor CR. Antigen retrieval immunohistochemistry: Past, present and future. J Histochem Cytochem 1997;45:327-43. |
|38.||Al-Abadi H, Nagel R, Neuhaus P. Immunohistochemical detection of p53 protein in transitional cell carcinoma of the bladder in correlation to DNA ploidy and pathohistological stage and grade. Cancer Detect Prev 1998;22:43-50. |
|39.||Lin HY, Huang CH, Wu WJ, Chou YH, Fan PL, Lung FW. Mutation of the p53 tumor suppressor gene in transitional cell carcinoma of the urinary tract in Taiwan. Kaohsiung J Med Sci 2005;21:57-64. |
|40.||Moch H, Sauter G, Moore D, Mihatsch MJ, Gudat F, Waldman F. p53 and erbB-2 protein over expression are associated with early invasion and metastasis in bladder cancer. Virchows Arch A Pathol Anat Histopathol 1993;423:329-34. |
|41.||Kuczyk MA, Serth J, Hervatin C, Arndt H, Derendorf L, Thon WF, et al . Detection of p53 tumor-suppressor-gene protein in bladder tumors and prostate cancer: Possible clinical implications. World J Urol 1994;12:345-51. |
|42.||Kong G, Shin KY, Oh YH, Lee JJ, Park HY, Woo YN, et al . Bcl-2 and p53 expressions in invasive bladder cancers. Acta Oncol 1998;37:715-20. |
|43.||Underwood MA, Reeves J, Smith G, Gardiner DS, Scott R, Bartlett J, et al . Overexpression of p53 protein and its significance for recurrent progressive bladder tumours. Br J Urol 1996;77:659-66. |
|44.||Okamura T, Umemoto Y, Yasui T, Saiki S, Kuroda H, Kotoh S, et al . Noninvasive detection of alterations in chromosome numbers in urinary bladder cancer cells, using fluorescence in situ hybridization. Int J Clin Oncol 2004;9:373-7. |
|45.||Li B, Kanamaru H, Noriki S, Fukuda M, Okada K. Numeric aberration of chromosome 17 is strongly correlated with p53 overexpression, tumor proliferation and histopathology in human bladder cancer. Int J Urol 1998;5:317-23. |
|46.||Catalona WJ. Urothelial tumors of the urinary tract. In: Walsh PC, Retik AB, Stamey TA, Vaughan ED, editors. Campbell's urology. 5 th ed. Philadelphia: Saunders Co; 1992. p. 2285-353. |
Clinical Pharmacy Laboratory, Drug Applied Research Center, Pashmineh, Daneshgah Street, Tabriz
Source of Support: None, Conflict of Interest: None
[Figure 1a], [Figure 1b], [Figure 2], [Figure 3]
|This article has been cited by|
||Neoplasias uroteliais papilíferas superficiais da bexiga (pTa e pT1): correlação da expressão do p53, KI-67 E CK20 com grau histológico, recidiva e progressão tumoral
| ||Daniel Cury Ogata,Carmen Austrália Paredes Ribas Marcondes,Felipe Francisco Tuon,Wilson Francisco Schreiner Busato Júnior,Gabriela Cavalli,Letícia Elizabeth Augustin Czeczko |
| ||Revista do Colégio Brasileiro de Cirurgiões. 2012; 39(5): 394 |
|[Pubmed] | [DOI]|
||Spotlight on Differentially Expressed Genes in Urinary Bladder Cancer
| ||Apostolos Zaravinos,George I. Lambrou,Dimitrios Volanis,Dimitris Delakas,Demetrios A. Spandidos,I. King Jordan |
| ||PLoS ONE. 2011; 6(4): e18255 |
|[Pubmed] | [DOI]|
| Article Access Statistics|
| Viewed||4051 |
| Printed||105 |
| Emailed||1 |
| PDF Downloaded||378 |
| Comments ||[Add] |
| Cited by others ||2 |