Comparative quantitative evaluation of the XIAP, survivin & Ki67 transcript levels in urine & tissue...

Comparative quantitative evaluation of the XIAP, survivin & Ki67 transcript levels in urine & tissue

samples of bladder cancer patients

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

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

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

Materials & Methods

• Exclusion criteria

– Patients with PCa, IDC, UTI

• Controls

– BPH patients

– Cystitis patients

– Healthy volunteers

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

Materials & Methods

• Quantitative evaluation of urinary & tissue

levels of survivin, XIAP & Ki67

• Normalized ratios of survivin, XIAP &

Ki67 correlated with clinicopathological

data

Results

• Reference genes used – HPRT & TBP

• Negative correlation between reference gene

expression & urinary contamination by RBCs &

WBCs

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)

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%

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%

benign atypia low high NA

9 (15.3%) 2 (3.3%) 9 (15.3%) 18 (30.5%) 21 (35.6%)

BCa patients

• 59 primary TURBT

42 second op (33 sec TUR, 9 cystec)

8 third op (6 tertiary TUR, 2 cystec)

Controls

BPH Cystitis Healthy

53 13 27

68 yrs (52-85) 64 yrs (19-85) 31 yrs (18-59)

PSA median 2.9 (0.3 – 33.9)

M : F3 (23.1%) :10 (76.9%)

M : F7 (25.9 %) :20 (74.1 %)

Tumor markers between BCa patients & controls

Clinical diagnosis

Ki67/

HPRT

Ki67/

TBP

XIAP/

HPRT

XIAP/

TBP

SVV/

HPRT

SVV/

TBP

Bca (48)(positive histo)

0.82 1.96 28.33 47.25 1.50 2.59

No tumor (11)(negative histo)

0.52 1.73 14.74 37.65 1.11 1.23

BPH (53) 0.40 1.02 19.29 45.65 1.08 2.29

Cystitis (13) 0.79 0.31 73.98 42.44 1.35 0.56

Healthy (27) 0.00 0.00 39.81 17.32 2.71 3.14

Median values presented

Ki67

Verarbeitete Fälle

34 70,8% 14 29,2% 48 100,0%

11 100,0% 0 ,0% 11 100,0%

41 77,4% 12 22,6% 53 100,0%

12 92,3% 1 7,7% 13 100,0%

12 44,4% 15 55,6% 27 100,0%

UrineBT

NT

BPH

Cystitis

Healthy

Ki67/HPRT

N Prozent N Prozent N Prozent

Gültig Fehlend Gesamt

Fälle

BT NT BPH Cystitis Healthy

Urine

0

1

2

3

4

5

6

Ki6

7/H

PR

T

52

98

66

67

29

53

100

81

76

103

125

146

Verarbeitete Fälle

36 75,0% 12 25,0% 48 100,0%

11 100,0% 0 ,0% 11 100,0%

44 83,0% 9 17,0% 53 100,0%

13 100,0% 0 ,0% 13 100,0%

16 59,3% 11 40,7% 27 100,0%

UrineBT

NT

BPH

Cystitis

Healthy

Ki67/TBP



Fälle


Urine

0

2

4

6

8

10

Ki6

7/T

BP

20

40

53

100

43

10

118

93

103

81

98

125

76

67

60

XIAP

Verarbeitete Fälle

34 70,8% 14 29,2% 48 100,0%

11 100,0% 0 ,0% 11 100,0%

41 77,4% 12 22,6% 53 100,0%

12 92,3% 1 7,7% 13 100,0%

9 33,3% 18 66,7% 27 100,0%

UrineBT

NT

BPH

Cystitis

Healthy

XIAP/HPRT



Fälle


Urine

0

25

50

75

100

125

XIA

P/H

PR

T

39

17 53

99

23

41

46

57

125

79

74

92

86

69

63

Verarbeitete Fälle

36 75,0% 12 25,0% 48 100,0%

11 100,0% 0 ,0% 11 100,0%

44 83,0% 9 17,0% 53 100,0%

13 100,0% 0 ,0% 13 100,0%

10 37,0% 17 63,0% 27 100,0%

UrineBT

NT

BPH

Cystitis

Healthy

XIAP/TBP



Fälle


Urine

0

100

200

300

XIA

P/T

BP

23

17

121

7

25

41

57

99

74

125

79

92

104

86

94

62

63

69

Survivin

Verarbeitete Fälle

26 54,2% 22 45,8% 48 100,0%

8 72,7% 3 27,3% 11 100,0%

27 50,9% 26 49,1% 53 100,0%

5 38,5% 8 61,5% 13 100,0%

10 37,0% 17 63,0% 27 100,0%

UrineBT

NT

BPH

Cystitis

Healthy

SVV/HPRT



Fälle


Urine

0

2

4

6

8

10

12

SV

V/H

PR

T

103

130

29

46

74

125

Verarbeitete Fälle

27 56,3% 21 43,8% 48 100,0%

8 72,7% 3 27,3% 11 100,0%

26 49,1% 27 50,9% 53 100,0%

5 38,5% 8 61,5% 13 100,0%

14 51,9% 13 48,1% 27 100,0%

UrineBT

NT

BPH

Cystitis

Healthy

SVV/TBP



Fälle


Urine

0

2

4

6

8

10

12

SV

V/T

BP

29

25

93

74

125

133

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)

0.52 1.73 14.74 37.65 1.11 1.23

Superficial (38)

0.82 2.05 22.02 50.21 1.48 2.65

Invasive (10)

1.79 2.84 48.9 63.35 1.51 1.64


Ki67

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

25 65,8% 13 34,2% 38 100,0%

9 90,0% 1 10,0% 10 100,0%

BT stageNT

superficial

invasive

Ki67/HPRT



Fälle

NT superficial invasive

Bt stage

0

1

2

3

4

5

6

Ki6

7 /

HP

RT

4

5

34

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

27 71,1% 11 28,9% 38 100,0%

9 90,0% 1 10,0% 10 100,0%

BT stageNT

superficial

invasive

Ki67/TBPN Prozent N Prozent N Prozent


Fälle


Bt stage

0

3

6

9

12

15

Ki6

7 /

TB

P

5

43

46

34 52

XIAP

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

25 65,8% 13 34,2% 38 100,0%

9 90,0% 1 10,0% 10 100,0%

BT stageNT

superficial

invasive

XIAP/HPRT



Fälle


Bt stage

0

50

100

150

200

XIA

P /

HP

RT

5

42

9

23

44

58

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

27 71,1% 11 28,9% 38 100,0%

9 90,0% 1 10,0% 10 100,0%

BT stageNT

superficial

invasive

XIAP/TBP



Fälle


BT stage

0

100

200

300

400

XIA

P /

TB

P

9

23

16

30

44

Survivin

Verarbeitete Fälle

8 72,7% 3 27,3% 11 100,0%

19 50,0% 19 50,0% 38 100,0%

7 70,0% 3 30,0% 10 100,0%

BT stageNT

superficial

invasive

SVV/HPRT



Fälle


BT stage

0

1

2

3

4

SV

V /

HP

RT

34

58

Verarbeitete Fälle

8 72,7% 3 27,3% 11 100,0%

20 52,6% 18 47,4% 38 100,0%

7 70,0% 3 30,0% 10 100,0%

BT stageNT

superficial

invasive

SVV/TBPN Prozent N Prozent N Prozent


Fälle


BT stage

0

2

4

6

8

10

12

SV

V /

TB

P

9 43

46

34

30

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)

0.52 1.73 14.74 37.65 1.11 1.23

Low grade (14)

0.60 0.73 33.60 38.19 1.61 2.70

High grade (34)

0.91 2.18 23.35 81.29 1.20 2.34


Ki67

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

11 73,3% 4 26,7% 15 100,0%

23 69,7% 10 30,3% 33 100,0%

BT gradeNT

low

high

Ki67/HPRT



Fälle

NT low high

BT grade

0

1

2

3

4

5

6

Ki6

7 /

HP

RT

4

48

5

40

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

11 73,3% 4 26,7% 15 100,0%

25 75,8% 8 24,2% 33 100,0%

BT gradeNT

low

high



Fälle

NT low high

BT grade

0

5

10

15

Ki6

7 /

TB

P

5

23

48

53

XIAP

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

11 73,3% 4 26,7% 15 100,0%

23 69,7% 10 30,3% 33 100,0%

BT gradeNT

low

high

XIAP /HPRT



Fälle

NT low high

Bt grade

0

25

50

75

100

125

XIA

P /

HP

RT

5 32

56

9

24

26

Verarbeitete Fälle

11 100,0% 0 ,0% 11 100,0%

11 73,3% 4 26,7% 15 100,0%

25 75,8% 8 24,2% 33 100,0%

BT gradeNT

low

high

XIAP/TBP



Fälle

NT low high

BT grade

0

100

200

300

400

500

XIA

P /

TB

P

9

24

28

37

Survivin

Verarbeitete Fälle

8 72,7% 3 27,3% 11 100,0%

6 40,0% 9 60,0% 15 100,0%

20 60,6% 13 39,4% 33 100,0%

BT gradeNT

low

high

SVV/HPRT



Fälle

NT low high

BT grade

0

1

2

3

4

SV

V /

HP

RT

26

40

Verarbeitete Fälle

8 72,7% 3 27,3% 11 100,0%

5 33,3% 10 66,7% 15 100,0%

22 66,7% 11 33,3% 33 100,0%

BT gradeNT

low

high



Fälle

NT low high

BT grade

0

5

10

15

20

SV

V /

TB

P

9

14

12

40

37

Tumor markers between first & second TURBT

BT paired urine samples (33)

Ki67/

HPRT

Ki67/

TBP

XIAP/

HPRT

XIAP/

TBP

SVV/

HPRT

SVV/

TBP

Op1 0.69 1.81 21.63 55.95 1.34 2.57

Op2 0 0 25.19 83.45 0.95 2.25


Ki67

Verarbeitete Fälle

23 69,7% 10 30,3% 33 100,0%

22 66,7% 11 33,3% 33 100,0%

Urineprimäre TURBT

sekundäreTURBT

Ki67/HPRTN Prozent N Prozent N Prozent


Fälle

primäre TURBT sekundäre TURBT

Urine

0,0

0,5

1,0

1,5

2,0

2,5

Ki6

7 /

HP

RT

28

31

21

43

60

39

Verarbeitete Fälle

25 75,8% 8 24,2% 33 100,0%

23 69,7% 10 30,3% 33 100,0%

Urineprimäre TURBT

sekudäre TURBT



Fälle


Urine

0

1

2

3

4

Ki6

7 /

TB

P

27

28

31

21

43

60

39

XIAP

Verarbeitete Fälle

23 69,7% 10 30,3% 33 100,0%

22 66,7% 11 33,3% 33 100,0%

Urineprimäre TURBT

sekundäreTURBT

XIAP/HPRTN Prozent N Prozent N Prozent


Fälle


Urine

0

50

100

150

200

250

300

XIA

P /

HP

RT

11

55

46

30

61

41

Verarbeitete Fälle

25 75,8% 8 24,2% 33 100,0%

23 69,7% 10 30,3% 33 100,0%

Urineprimäre TURBT

sekundäreTURBT

XIAP/TBPN Prozent N Prozent N Prozent


Fälle


Urine

0

100

200

300

400

XIA

P /

TB

P

11

6

17

30

41

61

40

Survivin

Verarbeitete Fälle

19 57,6% 14 42,4% 33 100,0%

14 42,4% 19 57,6% 33 100,0%

Urineprimäre TURBT

sekundäreTURBT

SVV/HPRTN Prozent N Prozent N Prozent


Fälle


Urine

0,0

0,5

1,0

1,5

2,0

2,5

SV

V /

HP

RT

11

31

46

21

50

61

Verarbeitete Fälle

20 60,6% 13 39,4% 33 100,0%

14 42,4% 19 57,6% 33 100,0%

Urineprimäre TURBT

sekundäre TURBT

SVV/TBP



Fälle


Urine

0

2

4

6

8

SV

V /

TB

P

11

46

50

31

21

17

61

Tumor markers for unpaired tissue samples

tissue samples

Ki67/

HPRT

Ki67/

TBP

XIAP/

HPRT

XIAP/

TBP

SVV/

HPRT

SVV/

TBP

Tumor tissue (26)

1.55 5.08 6.13 13.98 0.41 1.03

Tumor free (27)

1.24 1.23 8.25 11.92 0.26 0.35


Ki67

Verarbeitete Fälle

26 100,0% 0 ,0% 26 100,0%

27 100,0% 0 ,0% 27 100,0%

HistologieTu

Tf

Ki67/HPRT



Fälle

Tf Tu

Histologie

0

1

2

3

4

5

6

Ki6

7 /

HP

RT

20

26

17

21

35

52

Verarbeitete Fälle

26 100,0% 0 ,0% 26 100,0%

27 100,0% 0 ,0% 27 100,0%

HistologieTu

Tf

Ki67/TBP



Fälle

Tf Tu

Histologie

0

3

6

9

12

15

Ki6

7 /

TB

P

26

17

5235

XIAP

Verarbeitete Fälle

26 100,0% 0 ,0% 26 100,0%

27 100,0% 0 ,0% 27 100,0%

HistologieTu

Tf

XIAP/HPRT



Fälle

Tf Tu

Histologie

0

5

10

15

20

XIA

P /

HP

RT

52

Verarbeitete Fälle

26 100,0% 0 ,0% 26 100,0%

27 100,0% 0 ,0% 27 100,0%

HistologieTu

Tf

XIAP/TBP



Fälle

Tf Tu

Histologie

0

5

10

15

20

25

30

XIA

P /

TB

P

14

2

32

17

52

Survivin

Verarbeitete Fälle

26 100,0% 0 ,0% 26 100,0%

27 100,0% 0 ,0% 27 100,0%

HistologieTu

Tf

SVV/HPRT



Fälle

Tf Tu

Histologie

0

0

1

2

2

SV

V /

HP

RT

53

21

17

45

52

35

Verarbeitete Fälle

26 100,0% 0 ,0% 26 100,0%

27 100,0% 0 ,0% 27 100,0%

HistologieTu

Tf

SVV/TBP



Fälle

Tf Tu

Histologie

0

1

2

3

4

5

SV

V /

TB

P

45

17

52

35

Tumor markers for paired tissue samples

Paired tissue samples (13)

Ki67/

HPRT

Ki67/

TBP

XIAP/

HPRT

XIAP/

TBP

SVV/

HPRT

SVV/

TBP

Tumor tissue

1,73 5,99 6,42 13,61 0,53 1,11

Tumor free

1,52 2,01 8,22 14,26 0,35 0,41


Ki67

Verarbeitete Fälle

13 100,0% 0 ,0% 13 100,0%

13 100,0% 0 ,0% 13 100,0%

TissueTf

Tu

Ki67/HPRT



Fälle

Tf Tu

Tissue

0

2

4

6

8

Ki6

7 /

HP

RT

10

16

Verarbeitete Fälle

13 100,0% 0 ,0% 13 100,0%

13 100,0% 0 ,0% 13 100,0%

TissueTf

Tu



Fälle

Tf Tu

Tissue

0

2

4

6

8

10

12

Ki6

7 /

TB

P

4

16

XIAP

Verarbeitete Fälle

13 100,0% 0 ,0% 13 100,0%

13 100,0% 0 ,0% 13 100,0%

TissueTf

Tu

XIAP/HPRT



Fälle

Tf Tf

Tissue

2

4

6

8

10

12

14

XIA

P /

HP

RT

15

Verarbeitete Fälle

13 100,0% 0 ,0% 13 100,0%

13 100,0% 0 ,0% 13 100,0%

TissueTf

Tu

XIAP/TBPN Prozent N Prozent N Prozent


Fälle

Tf Tu

Tissue

0

5

10

15

20

XIA

P /

TB

P

19

12

Survivin

Verarbeitete Fälle

13 100,0% 0 ,0% 13 100,0%

13 100,0% 0 ,0% 13 100,0%

TissueTf

Tu

SVV/HPRT



Fälle

Tf Tu

Tissue

0,0

0,5

1,0

1,5

2,0

SV

V /

HP

RT

21

16

10

Verarbeitete Fälle

13 100,0% 0 ,0% 13 100,0%

13 100,0% 0 ,0% 13 100,0%

TissueTf

Tu



Fälle

Tf Tu

Tissue

0

1

2

3

4

5

SV

V /

TB

P

21

16

Ratio Tu / Tf

Paired tissue samples (13)

Ki67/

HPRT

Ki67/

TBP

XIAP/

HPRT

XIAP/

TBP

SVV/

HPRT

SVV/

TBP

Median 3.32 3.39 0.72 0.71 1.67 2.66

>1 8 (61.5%)

10 (76.9%)

3 (23.1%)

6 (46.2%)

8 (61.5%)

8 (61.5%)

<1 5 (38.5%)

3 (23.1%)

10 (76.6%)

7 (53.8%)

5 (38.5%)

5 (38.5%)

Comparative quantitative evaluation of the XIAP, survivin & Ki67 transcript levels in urine & tissue...

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Transcript of Comparative quantitative evaluation of the XIAP, survivin & Ki67 transcript levels in urine & tissue...