1. Comparative quantitative evaluation of the XIAP, survivin & Ki67 transcript levels in urine & tissue samples of bladder cancer patients

2. Introduction Bladder cancer (BCa)- 5th commonest cancer in European men & 6th leading cause of death Cystoscopy & urine cytology - current gold standards for diagnosis & surveillance of BCa Ideal tumour marker for non-invasive diagnosis & surveillance does not yet exist

3. Introduction Survivin inhibitor of apoptosis protein (IAP) that is selectively over- expressed in most human malignancies Objectives – determine suitability of survivin as diagnostic, surveillance (and prognostic) marker of BCa –compare the utility of survivin to other potential tumour markers e.g. Ki67 and XIAP

4. Materials & Methods Prospective study Single, tertiary level referral centre January - June 2006 Inclusion criteria –patients undergoing transurethral resection (TUR) for newly diagnosed BCa, recurrent BCa & cystoscopically suspicious bladder lesions

5. Materials & Methods Exclusion criteria –Patients with PCa, IDC, UTI Controls –BPH patients –Cystitis patients –Healthy volunteers

6. Materials & Methods BCa patients –Pre-operative urine sample –Intra-operative tumour tissue & “normal appearing” bladder mucosa –Post-operative urine sample (1 POD) –Above repeated for every session of TUR Controls –1 urine sample from each

7. Materials & Methods Quantitative evaluation of urinary & tissue levels of survivin, XIAP & Ki67 Normalized ratios of survivin, XIAP & Ki67 correlated with clinicopathological data

8. Results Reference genes used – HPRT & TBP Negative correlation between reference gene expression & urinary contamination by RBCs & WBCs

9. BCa patients 65 recruited 6 pts with both BCa & Pca, excluded from analysis > 59 pts analysed Age (median)= 69yrs (34 – 89) M:F= 39: 20 (66.1% : 33.9%) Newly dx: BCa recurr= 54: 5 (91.5% : 8.5%) PSA (median; 36 pts)= 0.90 (0.16 – 12.54)

10. BCa patients Stage @ diagnosis (59) No tumour (11) 18.6% Superficial (38) = Ta+T1 64.5% Invasive (10) = T2 16.9% Grade @ diagnosis (59) No tumour (11) 18.6% Low (14) = G1 23.8% High (34) = G2+G3 57.6%

11. BCa patients cis –Pos: neg= 5: 54 (8.5% : 91.5%) –All with cis harbour high grade (G2/ G3) disease as well Cytology –Sensitivity= 65.2%; Specificity= 80.0% –PPV= 83.3%; NPV= 60.0% benignatypialowhighNA 9 (15.3%)2 (3.3%)9 (15.3%)18 (30.5%)21 (35.6%)

12. BCa patients 59 primary TURBT 42 second op (33 sec TUR, 9 cystec) 8 third op (6 tertiary TUR, 2 cystec)

13. Controls BPHCystitisHealthy 531327 68 yrs (52-85)64 yrs (19-85)31 yrs (18-59) PSA median 2.9 (0.3 – 33.9) M : F 3 (23.1%) :10 (76.9%) M : F 7 (25.9 %) :20 (74.1 %)

14. Tumor markers between BCa patients & controls Clinical diagnosis Ki67/ HPRT Ki67/ TBP XIAP/ HPRT XIAP/ TBP SVV/ HPRT SVV/ TBP Bca (48) (positive histo) 0.821.9628.3347.251.502.59 No tumor (11) (negative histo) 0.521.7314.7437.651.111.23 BPH (53) 0.401.0219.2945.651.082.29 Cystitis (13) 0.790.3173.9842.441.350.56 Healthy (27) 0.00 39.8117.322.713.14 Median values presented

15. Ki67

16. XIAP

17. Survivin

18. Tumor markers vs BCa stage BT stage @ pri TUR (59) Ki67/ HPRT Ki67/ TBP XIAP/ HPRT XIAP/ TBP SVV/ HPRT SVV/ TBP No tumor (11) 0.521.7314.7437.651.111.23 Superficial (38) 0.822.0522.0250.211.482.65 Invasive (10) 1.792.8448.963.351.511.64 Median values presented

19. Ki67

20. XIAP

21. Survivin

22. Tumor markers vs BCa grade BT grade @ pri TUR (59) Ki67/ HPRT Ki67/ TBP XIAP/ HPRT XIAP/ TBP SVV/ HPRT SVV/ TBP No tumor (11) 0.521.7314.7437.651.111.23 Low grade (14) 0.600.7333.6038.191.612.70 High grade (34) 0.912.1823.3581.291.202.34 Median values presented

23. Ki67

24. XIAP

25. Survivin

26. Tumor markers between first & second TURBT BT paired urine samples (33) Ki67/ HPRT Ki67/ TBP XIAP/ HPRT XIAP/ TBP SVV/ HPRT SVV/ TBP Op1 0.691.8121.6355.951.342.57 Op2 0025.1983.450.952.25 Median values presented

27. Ki67

28. XIAP

29. Survivin

30. Tumor markers for unpaired tissue samples tissue samples Ki67/ HPRT Ki67/ TBP XIAP/ HPRT XIAP/ TBP SVV/ HPRT SVV/ TBP Tumor tissue (26) 1.555.086.1313.980.411.03 Tumor free (27) 1.241.238.2511.920.260.35 Median values presented

31. Ki67

32. XIAP

33. Survivin

34. Tumor markers for paired tissue samples Paired tissue samples (13) Ki67/ HPRT Ki67/ TBP XIAP/ HPRT XIAP/ TBP SVV/ HPRT SVV/ TBP Tumor tissue 1,735,996,4213,610,531,11 Tumor free 1,522,018,2214,260,350,41 Median values presented

35. Ki67

36. XIAP

37. Survivin

38. Ratio Tu / Tf Paired tissue samples (13) Ki67/ HPRT Ki67/ TBP XIAP/ HPRT XIAP/ TBP SVV/ HPRT SVV/ TBP Median 3.323.390.720.711.672.66 >18 (61.5%) 10 (76.9%) 3 (23.1%) 6 (46.2%) 8 (61.5%) <15 (38.5%) 3 (23.1%) 10 (76.6%) 7 (53.8%) 5 (38.5%)