Research articles
 

By Mr. Anthony K Venyo , Mr. Harold Greenwood , Mr. Ian Dimmick , Mr. David W Herring , Dr. Douglas J Maloney
Corresponding Author Mr. Anthony K Venyo
Urology Department. North Manchester General Hospital, - United Kingdom M8 5RB
Submitting Author Mr. Anthony Kodzo - Grey Venyo
Other Authors Mr. Harold Greenwood
University Hospital of North Durham Department of Pathology, - United Kingdom

Mr. Ian Dimmick
University Hospital of North Durham Department of Haematology Durham United Kingdom, - United Kingdom

Mr. David W Herring
University Hospital of North Durham Department of Surgery, - United Kingdom

Dr. Douglas J Maloney
Airedale General Hospital Skipton Road Skeeton Keighley West Yorkshire BD20 6TW United Kingdom., - United Kingdom

UROLOGY

Urothelial carcinoma, DNA ploidy, Aneuploidy, p53, Immunohistochemistry, Flow cytometer, Over-expression, Cancer, Bladder

Venyo AK, Greenwood H, Dimmick I, Herring DW, Maloney DJ. DNA Ploidy And The Expression Of P53 In Human Urothelial Carcinoma. WebmedCentral UROLOGY 2011;2(1):WMC001461
doi: 10.9754/journal.wmc.2011.001461
No
Submitted on: 18 Jan 2011 01:09:41 PM GMT
Published on: 18 Jan 2011 08:31:14 PM GMT

Abstract


Background
There is no clear cut way to predict exactly which superficial urothelial carcinomas would subsequently not recur, which tumours would subsequently recur or progress after initial resection of the tumours. There is also no accurate way of predicting the outcome of muscle-invasive tumours urothelial following treatment
Aims
(a) To confirm the suggestion that aneuploidy is more common in urothelial tumours of high grade, high stage and is associated with inferior prognosis.
(b) To find out if the combination of aneuploidy and over-expression of p53 would be more selective in predicting superficial tumours that would progress and in predicting muscle-invasive tumours of poor prognosis.
Methods
Correlation of the DNA ploidy state and the expression of p53 in human urothelial carcinoma was studied using flow cytometry and an immuno-histological (ABC) method. The ploidy state of 34 tumours were analysed with regards to grade, stage and outcome. The expression of p53 was also analysed with regards to grade, stage and outcome. Patients who had trans-urethral resection of bladder tumour had regular check cystoscopies with biopsies / resection of any recurrent tumours. Patients with muscle-invasive tumours who were treated by radiotherapy or cystectomy were reviewed regularly in the outpatient clinic and were assessed by clinical examination and ultrasound scan / CT scan of the abdomen pelvis and chest as may be required. They also had routine blood tests including: Full blood count; Serum urea and electrolytes; and liver function tests.
Results
Three out of 12 G1, 6 out of 9 G2 and 8 out of 12 G3 tumours respectively were aneuploid. Nine out of 12 G1, 3 out of 9 G2 and 4 out of 12 G3 tumours respectively were diploid. These results confirm the suggestion that aneuploidy is more commonly associated with urothelial carcinomas of high grade and high stage. All the 3 superficial tumours that progressed were aneuploid. However, the ploidy state of the tumours did not appear to have influenced the outcome of the muscle-invasive tumours. All the three superficial tumours that progressed were also observed to be moderately or strongly positively stained for p53. The three superficial tumours that progressed were therefore aneuploid and p53 positive. Aneuploidy and over-expression of p53 did not appear to have influenced the outcome of the muscle-invasive tumours.
Conclusions
These results confirm the suggestion that aneuploidy is more commonly associated with urothelial carcinomas of high grade and high stage.
All the three Superficial tumours that progressed were aneuploid.
All the three superficial tumours that progressed had over-expression of p53 (were moderately or strongly positive for p53)
All the three superficial tumours that progressed were aneuploid as well as had over-expression of p53
Aneuploidy and over-expression of p53 did not appear to have influenced the outcome of muscle-invasive tumours.


Introduction


It is believed that diploid urothelial tumours generally tend to be low-grade and low-stage and patients with diploid tumours are said to have a favourable prognosis. Tumours with triploid to tetraploid chromosome number are believed to have unfavourable prognosis. It has been suggested that patients with tumours that are tetraploid or have a greater than tetraploid chromosome number have a more favourable prognosis than triploid to tetraploid tumours but a worse prognosis than patients with diploid tumours [1], [2] [3].. An additional suggestion is that for predicting the likelihood of tumour progression, flow cytometry is limited in view of the fact that a significant proportion of bladder cancers that progress are diploid or near diploid and some superficial tumours that do not progress are aneuploid [4].
Lipponen [5], studied 212 archival paraffin-embedded samples of bladder cancer using CMI polyclonal antibody. In this study 29% of tumours were stained positively for p53 protein and over-expression was stated to be associated with of tumour progression, histological grade, non-papillary growth architecture, dense inflammatory reaction, DNA aneuploidy, high S-phase fraction and mitotic frequency. Over-expression of p53 was found to predict poor outcome in muscle-invasive but not in superficial tumours.
This study was carried out in order to : (a) confirm the suggestion that aneuploidy is more common in urothelial tumours of high grade and high stage and is associated with inferior prognosis. (b) find out if the combination of aneuploidy and over-expression of p53 would be more selective in predicting superficial tumours that would progress and in predicting muscle-invasive tumours of poor prognosis.

Patients, Materials and Methods.


Fresh samples of urothelial carcinoma were obtained from 34 patients by transurethral resection or radical excision (cystectomy). The specimens were stored in liquid nitrogen. Tumour grade was carried out by the pathologist. Tumour stage was assessed using standard clinical, radiological and histological criteria. Immunohistochemistry for p53 was carried out using the sections of fresh urothelial cancer specimens, standard immunohistochemical staining methods and primary antisera Novo Castria NCL-DO7 anti p53. Staining was assessed taking into consideration the intensity of positive staining through out the section. Staining intensity was scored on a 4 point scale. Negative (no stain) 0, weak (1), moderate (2) and strong (3). The extent of staining was based upon the proportion of tumour cells positively stained. 0-25% +, 25-50% ++, 50-75% +++, and 75-100% ++++. In the final analysis of data all tumours showing ≥ 10% weak, moderate and strong staining were recorded as positive (see illustrations 7 and 8 [figures 1 and 2] which illustrate strong and moderate staining for p53). For DNA ploidy analysis, standard flow cytometry (see illustration 9 [figure 3] which illustrates the flow cytometer which was used for the study) was performed using (a) fresh normal venous blood samples obtained in EDTA and processed on a coulter DNA prep work station as normal control and (b) fresh urothelial carcinoma samples of 30 of the patients which were macerated in weigh boat and the cell count adjusted to 3-10 109/L using PBS PH7.2. The samples were processed against normal control on coulter Epics profile 11 and a plot of the ploidy state obtained against the normal diploid control.

Results:


Tumours from 34 patients were analysed by flow cytometry for DNA ploidy. There was not enough tumour for immuno-histochemistry for 4 patients. Nine out of 12 (75%) well differentiated (G1) tumours were diploid (see illustration 1 (table 1); illustration 10 (figure 4) illustrates a diploid tracing); 3 out of 9 (33.3%) of the moderately differentiated (G2) tumours were diploid; 4 out of 12 (33.3%) poorly differentiated (G3) tumours were diploid.
The results of the study would suggest that aneuploid tumours are more likely to be high grade in that 3 out of 12 G1 tumours (25%), 6 out of 9 (66.7%) of G2 tumours and 8 out of 12 (66.7%) G3 tumours were aneuploid respectively (see illustration1; i;;ustration 11 shows an aneuploid tracing).
With regard to stage of tumours, 9 out of 23 superficial tumours (39%) and 9 out of 11 (82%) of muscle-invasive tumours were aneuploid (see illustration 1).
With regard to outcome, all the three superficial tumours that progressed to higher stage were aneuploid and no diploid tumour progressed (see illustration 2). On the other hand out of 10 patients with aneuploid muscle-invasive tumours 6 (60%) were alive and well without any evidence of tumour. ( 2 of these were originally superficial tumours that subsequently progressed). All the patients with diploid tumours died, 3 as a result of their tumours and one died of an unrelated cause.
The distribution of the tumours with regard to histological grade and expression of p53 weree as follows: 5 out of 10 (50%) well differentiated (Grade 1)tumours were p53 positive and 5 were p53 negative; 7 (77.7%) out of 9 moderately differentiated (Grade 2) tumours were p53 positive and 2 p53 negatve; 8 (72.7%) out of 11 poorly differentiated (Grade 3) tumours were p53 positive and 3 were p53 negatve (see illustration 3).
With regard to stage of tumours and expression of p53, 15 (75%) out of 20 superficial tumours were p53 positive and 5 were p53 negative. Of the 10 muscle-invasive tumours surprisingly 5 (50%) were p53 positive and 5 were p53 negative (see illustration 3).
Regarding the expression of p53 and outcome of superficial tumours, out of 13 superficial tumours that were postively stiained for p53 8 (61.5%) either did not recur or the recurrent tumour was of the same grade and stage (regarded as not progressed); one medium/strongly positively stained well differentiated (G1 tumour) recurred into a poorly differentiated (G3) tumour of the same histological stage (regarded in this study as progresed to a higher grade); 3 medium/strongly p53 positively stained tumours progressed to higher stage; 1 patient whose superficial tumour was medium/strongly positively stained or showing over-expression of p53 died of unrelated cause and therefore there was no chance to know whether this tumour would have progressed or not. All the 3 superficial tumours that progressed to a higher stage were moderately to strongly positive for p53. None of the 3 superficial tumours that were p53 negatively stained recurred (see illustration 4). On the other hand, with regard to the muscle-invasive tumours expression of p53 did not appear to be related to outcome. There were 7 muscle-invasive p53 positive tumours, (1 weakly positively stained and 6 with over-expression of p53 (medium to strong postive staning). Out of the 7 patients with p53 positive muscle-invasive tumours, 4 were alive and well with no evidence of residual tumour (one patient’s tumour was weakly positively stained and 3 patients had tumours with over-expression of p53); 2 patients with tumours over-expressing p53 (medium /strongly positively stained tumours) died as a result of their tumours; one patient whose tumour had over-expression of p53 died of an unrelated cause (see illustration 4). There were 6 patients with p53 negative muscle-invasive tumours and of these, 2 patients were alve and well without any evidence of tumour; the tumour progressed in 1 patient; and 3 patients died as a result of their tumours (see illustration 4).
The characteristics of the 30 tumours studied by immunohistochemical staining for p53 and flow cytometry for DNA ploidy were as follows:
11 were aneuploid (non diploid /hyper diplod) and p53 positive
5 were aneuploid (non diploid / hyper diploid) and p53 negative
9 were diploid and p53 positive
5 were diploid and p53 negative (see illustration 5).
The charatericstics of the tumours with regard to stage and grade were as follows:
Out of 20 superficial tumours, 6 were aneuploid and p53 positive; 2 were aneuploid and p53 negative; 9 were diploid and p53 positive; 3 diploid and p53 negative.
Out of 10 muscle-invasive tumours studied, 5 were aneuploid and p53 positive; 3 were aneuploid and p53 negative; 2 were diploid and p53 negative; there was no tumour found that was diploid and p53 positive.
Out of 10 well differentiated (Grade 1) tumours, none was aneuploid and p53 positive; 2 were aneuploid and p53 negative; 5 were dplod and p53 postve; 3 were diplod and p53 negatve.
Out of 9 moderately dfferentiated (Grade 2) tumours, 5 aneuploid and p53 postve; 1 was aneuploid and p53 negative; 2 were diploid and p53 postive; 1 was dploid and p53 negative.
Out of 11 poorly differentiated (Grade 3) tumours, 6 were aneuploid and p53 postive; 2 were aneuploid and p53 negative; 2 were diploid and p53 positive; 1 was diploid and p53 negative (see illustraton 5).
The outcome of the superficial tumours as well as muscle-nvasive tumours were assessed with regard to correlaton of their staining characterstics and DNA ploidy. The findings were as follows:
With regard to the outcome of the superfcial tumours,
1. out of 7 superfcal tumours that were aneuploid and p53 postive, 3 tumours (1 aneuploid and weakly p53 postive and 2 aneuploid with over-expresson of p53 [moderate / strong stanng for p53] did not progress in that there was either no recurrence of tumour or the recurrence was of the same grade and stage; 3 tumours that were aneuploid with over-expression of p53 (moderate / strongly postive for p53) had recurrences of higher stage and were adjudged to progressed to a higher stage; 1 patient wth an aneuploid tumour and over-expression of p53 died of an unrelated cause.
2. Out of 5 tumours that were diploid with over-expression of p53 (moderate / strongly postive staining for p53), 4 either did not recur or the recurrence was of the same grade and staged and was considered not to have progressed in grade or stage; 1 well differentiated (Grade 1) tumour recurred and the recurrent tumour was of the same stage but poorly differentiated (Grade 3) and was adjudged to have progressed in grade from grade 1 to grade 3.
3.diploid and weakly p53 positive tumour did not recur.
4.None of the 3 diploid and p53 negative tumours recurred (see illustration 6).
With regard to the outcome of muscle-invasive tumours,
out of 6 patients with muscle-invasive tumours that were aneuploid and positively stained for p53 (1 aneuploid and weakly positively stained for p53 and 5 aneuploid with over-expression of p53 (moderate / strongly positively stained for p53), 4 patients were alive and well with no evidence of tumour and 2 patients died as a result of their tumours (both had aneuploid tmours with over-expression of p53).
Out of 4 patients with aneuploid and p53 negative tumours, 2 patients were alive and well with no evidence of tumour; the tumour progressed in 1 patient; 1 patient died as a result of the tumour.
1 patient with a diploid tumour with over-expression of p53 (moderate / strong staining for p53) died of unrelated cause.
2 patients with diploid and p53 negative tumours died as a result of their tumours (see illustration 6).
Regarding the simultaneous use of two markers, all the 3 patients with superficial tumours that progressed were aneuploid as well as moderate to strongly positive for p53. Recurrence of higher grade but the same stage was noticed to have occurred in a tumour that was diploid and moderate to strongly positive for p53. On the other hand progression to a higher stage was not observed in tumours that were diploid and p53 negative. In the case of the muscle invasive tumours the marker status did not appear to be factor in the outcome. Four patients with aneuploid tumours that were also positive for p53 were alive and well and 2 patients with characteristically similar tumours died as a result of their tumours. Two patients with p53 negative and diploid tumours died as a result of their tumours (see illustration 6).

Discussion


The recurrence rate of all newly diagnosed superficial carcinoma of the urinary bladder (pTa and pT1 tumours) is 50%. Most of the recurrences of superficial (pTa and pT1) tumours are of the same stage. About 15% of newly diagnosed carcinoma of the bladder subsequently progress. 60% of patients with muscle-invasive cancer of the urinary bladder die following treatment. There is no clear cut way of predicting which superficial urothelial carcinomas would subsequently progress and of predicting which muscle invasive tumours would be associated with death of the patient following treatment. It has been observed that the individual expression of (a) c-erb-B2 [6], (b) β-HCG [7], and (c) p53 [8] by superficial urothelial carcinomas may be associated with tumour recurrence and recurrence of high stage. It has also been observed that muscle-invasive tumours that are positively stained for (a) β-HCG [7] and (b) c-erb-B2 [6] may be of worse outcome in comparison to tumours that are negatively stained for any of the aforementioned markers. It has been noticed that urothelial tumours doubly positive for (a) β-HCG and c-erb-B2, (b) β-HCG and p53 and (c) c-erb-B2 and p53 would appear to have a worse outcome in comparison with tumours doubly negative for the aforementioned markers. However it was also observed that there is no statistical difference between the outcome of tumours that are doubly positive for two markers and tumours that are positive for one marker and negative for the second marker [9]. The results of this study confirm the suggestion that aneuploidy is associated with tumours of high stage and high grade. The results of this study may also possibly suggest that superficial tumours that progress may be aneuploid and positively stained for p53. The fact that not all the aneuploid superficial tumours and not all the p53 positive tumours progressed may suggest that the individual use of one of these two markers may not allow one the prediction of individual tumour progression. The finding in this study that progression of tumour was observed only in tumours that were aneuploid and p53 positive may initially appear to suggest that tumours that are aneuploid and p53 positive may be the tumours that would progress. The draw back in this suggestion is the fact that three tumours that were aneuploid and p53 positive did not progress. It may be interesting to notice that none of the tumours that were diploid and p53 negative progressed but the number of tumours involved in this study are small for any categorical emphatic comment to be made. The outcome of patients with muscle-invasive tumours would suggest that the simultaneous use of DNA ploidy and expression of p53 would not be of any benefit in predicting muscle invasive tumours that would have good or poor prognosis. Finally the distribution pattern of the tumour characteristics and the numbers involved in each group are too small to allow for any meaningful statistical analysis of the outcome of the various groups of tumours based upom their DNA ploidy state and p53 characteristics.
de Vere White and associates [10] evaluated the ability of DNA ploidy analysis to predict outcome of 228 patients with Ta/T1 transitional cell carcinomas (TCC). All patients were judged to be at increased risk for tumour recurrence due to having two occurences of stage T1 tumour within 56 weeks, or three or more tumours presenting simultaneously within 16 weeks of registration. Concurrent carcinoma in situ was acceptable. All patients were treated with either bacillus Calmette Guerin (BCG) immunotherapy or mitomycin C (MMC) intravesical chemotherapy. Patients with nondiploid tumours had higher hazard rates for both tumour progression and death (p = 0.007 and p = 0.016, respectively); however, the prognostic information of DNA ploidy was not additive to tumour grade.
Fossa and associates [11], studied DNA ploidy and S-phase fraction (SPF) determined by flow cytometry in 118 patients with muscle-invasive transitional cell carcinoma (TCC) of the urinary bladder scheduled for cystectomy after pre-operative radiotherapy (20 Gy/1 week) with or without systemic cisplatinin-based neo-adjuvant chemotherapy. The correlation between these parameters and immunohistochemically demonstrated p53, c-erbB-2 and HCG was additionally investigated. There were 102 DNA non-diploid tumours and 16 DNA diploid tumours. Fossa and associates [11], reported that DNA ploidy was not related to the T (all 118 patients) or pN (58 patients) category, occurence of stage reduction or cancer-related 5 years survival. Patients with high SPF tumours tended, however, to have a better prognosis in comparison with those with low SPF TCC reaching the level of significance (p < 0.05) for those patients who had high SPF tumours and received neo-adjuvant chemotherapy. Fifty-one of the tumours were p53 positive. P53 positive tumours were significantly more often found in TCC with low SPFs than in those with high SPFs. Respectively 12 and 9% of the tumours were HCG and c-erbB-2 positive, without correlation to DNA ploidy or SPF.
They concluded that:
DNA ploidy does not represent a prognostic parameter in muscle-invasive operable carcinomas.
A high SPF, determined by flowcytometry (FCM) may be helpful to identify patients with chemotherapy-sensitive TCC of the urinary bladder. .
Masters and associates [12] linked the histopathological grade, proportion of “S”-phase nuclei and DNA ploidy values and prognostic significance in a retrospective series of pT1 bladder cancers. They extracted nuclei from paraffin sections of 75 biopsies (56 patients). They used flow cytometry to measure DNA ploidy and the proportion of “S”-phase nuclei. They found that progressive disease (pT2 or greater) developed within 3 years in 35% (6/17) of patients with poorly differentiated tumours, 35% (8/23) with aneuploid tumours and 35% (7/20) of those with a high proportion “S”-phase nuclei. Of 8 tumours with all 3 features, progressive disease developed in 6 cases (75%). Of 9 patients who developed progressive disease, 8 (89%) had aneuploid tumours. Progressive disease did not develop in 11 patients with well differentiated tumours, in comparison with 4% (1/24) in diploid/tetraploid tumours and 7% (2/27) in those with a low/medium percentage of “S”-phase nuclei. In contrast to muscle-invasive disease, recurrent superficial tumours developed with a high incidence in all groups. Only 6/56 patients (11%) remained alive and disease-free for 3 years. Masters and associates concluded that these 3 features are of similar prognostic significance and accuracy in identifying patients requiring aggressive therapy.
Lipponen and associates [13] followed-up a cohort of 233 T2/T3 transitional cell carcinomas for over 10 years. Five nuclear factors, two mitotic indices, DNA ploidy and “S”-phase fraction (SPF) were related to progression and survival of transitional cell carcinomas (TCCs) during that time period. SPF predicted pelvic lymph node involvement at diagnosis (P = 0.064). Progression in T-category was related to T-category (P = 0.035), DNA ploidy (P = 0.0180), pappilary status (P = 0.0021), mitotic activity index (MAI) (P = 0.0011), volume corrected mitotic index (M/V index) (P = 0.0017), WHO grade (P = 0.0003), and “S”-phase fraction (P = 0.0002). Progression in N and M-categories were related to the same variables. Independent predictors of progression in T-category were SPF (P = 0.0161) and WHO grade (P = 0.00236), whereas, progression in M-category was independently related to MAI (P = 0.0012) and T-category (P = 0.0004). The SPF (P < 0.0001), M/V index (P < 0.0001), MAI (P < 0.0001), WHO grade (P < 0.0001) and papillary status (P < 0.0001) were the most important predictors of of survival in univariate analysis. In a multivariate analysis SPF and M/V index (P < 0.0001) were the best predictors of survival followed by papillary status and T-category. Liponen and associates [13] concluded that the results show that the proliferation rate of T2/T3 TCCs as determined by flow cytometric SPF or M/V index are equally powerful predictors; they (SPF and M/V index) are clearly better than nuclear morphometry, DNA ploidy or WHO grading as prognostic factors.
Loughman and associates [14] performed DNA image cytometry on 119 bladder biopsy supernate (BBS) specimens of transitional cell carcinoma (TCC) of the urinary bladder in order to test the suitability of this cytologic specimen for use in DNA ploidy analysis, and to assess the value of DNA ploidy measured on the specimen as to the risk of tumour recurrence and survival. Loughman and associates [14] correlated the histological grade and cytologic grade of the 119 tumours. The DNA ploidy produced was determined by image analysis of Feulgen-stained nuclei. They recorded the Kaplan-Meier curves related age, sex, grade, and DNA ploidy to recurrence of tumour and survival. Log rank analyses were used to ascertain the difference between the curves of eac categorical variable. Loughman and associates [14] stated that urothelial cells derived from BBS specimen were demontrated to be respresentative of the tumour. The tumour recurrence rate was significantly higher (P = 0.0001) and the survival rate significantly lower (P = 0.0002) for patients with aneuploid tumours compared in comparison with diploid tumours. Patients with 2 TCC tumours had significantly shorter time to recurrence (P = 0.003), although the relationship between ploidy and survival in this group was of marginal significance. Loughman and associates concluded that:
The specimen was free of many of the problems associated with the other types used for measuring DNA ploidy.
The results show that the BBS specimen is diagnostically useful for DNA analysis, providing prognostically relevant information.
Gransfors and associates [15] evaluated the predictive value of flow cytometry DNA analysis of cells from bladder washings in 43 patients with muscle-invasive or recurrent superficial bladder cancer treated with pre-operative radiotherapy and cystectomy. There was no correlation between ploidy status of the primary tumour and survival, neither was there any correlation between ploidy and the occurrence of residual tumour in the cystectomy specimens. Patients without residual tumours in the cystectomy specimen had significantly longer survival time than those with residual tumours. Granfors and associates [15] found that concomitant carcinoma in situ was correlated with better survival, which was surprising, considering the malignant potential of this lesion. Granfors and associates concluded that: in this study DNA ploidy did not predict tumour response to radiotherapy nor was it of any prognostic significance.
Lipponen and associates [16] measured by flow cytometry the nuclear DNA content of archival paraffin-embedded bladder cancer samples (70 patients) of WHO grades I-III. The female /male ratio was 15/55. The mean follow-up time was 13 years (range 9.6 -22.0 years). 37 of 70 (53%) patients had DNA index 1.0 (diploid DNA content), and the remaining 33 (47%) patients had aneuploid tumour. There was no significant difference in the age (mean ± SD) of the patients having a diploid (66 ± 9 years) or an aneuploid tumour (68 ± 11 years) at the time of diagnosis. There were 47 deaths during the follow-up period; 24 (51%) of these were as a result of bladder cancer (12 diploid, 12 aneuploid tumours). No significant difference was observed after radical treatment during the disease-free interval (mean ± SD) between diploid (48 ± 45 months) and aneuploid (35.5 ± 35 months) groups of patients. It was also observered that the recurrences during the follow-up period were equally common among diploid and aneuploid tumours. A statistically significant relation between histological grade and survival was demonstrated, however, DNA ploidy and “S”-phase fraction had little prognostic value in this respect. No statistically significant difference in survival was found between aneuploid (30%) and diploid (35%) groups of tumours during the follow-up period. Lipponen and associates [16] concluded that: The study suggests that flow cytometric determination of nuclear DNA ploidy from paraffin-embedded samples in bladder tumours does not add to the prognostic power of subjective histological grading.

Conclusions


These results confirm the suggestion that aneuploidy is more commonly associated with urothelial carcinomas of high grade and high stage.
All the three Superficial tumours that progressed were aneuploid.
All the three superficial tumours that progressed had over-expression of p53 (were moderately or strongly positive for p53)
All the three superficial tumours that progressed were aneuploid as well as had over-expression of p53
Aneuploidy and over-expression of p53 did not appear to have influenced the outcome of muscle-invasive tumours.

Acknowledgements


Acknowledgement to the following for allowing us to include some of their patients in the study:
i)  Mr J. Rhind, previously Consultant Urological Surgeon of Hartlepool General Hospital and currently Consultant Urological Surgeon in Dryburn Hospital, Durham.
ii)  Mr Trevor Armitage,  Consultant Urological Surgeon of South Shields General Hospital,  South Shields.
iii)  Mr B. S.  Fatya Vadanan, Consultant Urological Surgeon of Queen Elizabeth Hospital Gateshead.
iv)  Mr  Peter English, Consultant Urological Surgeon of Dryburn Hospital Durham.
v) Mr Ian Dimmick who was Chief  technician in the Haematology Department of Dryburn Hospital Durham City in the United Kingdom (this hospital is now called the University Hospital of North Durham North Road Durham City United kingdom) Mr Ian Dimmick was the expert on flow cytometry who told us there was a flow cytometer in the hospital which we could use. Mr Ian Dimmick showed us how the flow cytometer works and he supervised us on flow cytometry did the flow cytometry analysis. Mr Ian Dimmick was aware of the results of our study but subsequently moved to a new job and he is therefore a co-author of this paper. Without him we could not have made any progress with the flow cytometry.

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Ethical approval


Mr Anthony Kodzo-Grey Venyo designed the study and applied for and obtained ethical approval from the North Durham Ethics committee as well as obtained Research Registrar status from the Northern Regional Deanery in Newcastle upon Tyne in United Kingdom 

Source(s) of Funding


None

Competing Interests


None

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Consultant Radiologist
Posted by Dr. Emad Bakir on 21 Jan 2011 04:29:08 PM GMT

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