Supplementary Table 2: Non-proteomic studies to discover biomarkers of bladder
cancer
Biomarker(s)
‡:
MMP1
MMP2
MMP9
Samples†
n = 131 bladder tumour samples, n = 5
prostate cancer, n = 33 benign lower urinary
tract infections, n = 36 healthy volunteer.
n = 82 bladder carcinoma patients, n = 28
control patients.
n = 44 controls, n = 14 cystitis patients, n = 43
stage Ta-T1, n = 18 stage T2, n = 10 stage T3T4,
Gelatine zymography, Elevated in bladder
cancer and correlates with disease stage.
Sensitivity to detect bladder cancer = 0.51.
(3)
n = 82 bladder cancer patients (43 = pTa, 28
= pT1, 6 = pT2, 4 = pT3) and 28 controls.
ELISA and gelatin zymography. Elevated
MMP9 detects bladder cancer but with specificity
similar to urine cytology
(2)
n = 44 control urine samples, n = 14 cystitis
patients, n = 43 stage Ta-T1, n = 18 stage T2,
n = 10 stage T3-T4,
Gelatine zymography, Sensitivity to detect
bladder cancer = 0.31. MMP9 levels observed to
be elevated particularly within patients with
muscle invasive bladder cancer.
ELISA/zymography based study: Urinary
sediments used for cytology and within the
supernatant MMP2 : TIMP2 and MMP9 : TIMP2
ratios inversely correlated with tumour stage and
grade
Quantitative-WB: With optimal cut-off panel is
diagnostic of bladder cancer with 76.8%
sensitivity and 77.4% specificity.
(3)
Radioimmunoassay. EGF is decreased in
patients with bladder cancer (p < 0.0001).
(6)
Quantitative WB and ELISA: DEK protein
expressed in 33 out of 38 bladder tumours-not in
adjacent normal healthy tissues), and within 23
normal urine samples and 16 out of 19 bladder
cancer urine samples. 84% of transitional cell
carcinoma urine samples were tested positive for
the presence of DEK-protein.
ELISA test for cytokeratin 8/18 and UBC
ELISA test). Combination of the UBC antigen
test with the proportion of cells showing a DNA
content >2n increased the sensitivity of the UBC
test to 89%..
CYFRA21.1 test. At specificity of 95% cut-off
values for tests were: CYFRA21.1 = 5.4
micrograms/L, 15.4 micrograms/L for bladder
cancer antigen,760.8 units/L for tissue
polypeptide antigen and 14.6 U/ml for NMP22. At
these cut-offs the sensitivity of the CYFRA21.1
test for detection bladder cancer was 83.8%.
Immunoradiometric Assay. At 4.9
microgrammes/L CK19 fragments detect bladder
cancer with sensitivity = 79.3% and specificity =
88.6%.
(7)
Fluorescent antibody-based detectionImmunoCyt test: Multiple-antigen test detects
bladder cancer with 86.1% sensitivity and 79.4%
specificity.
(11)
HA-HAase test-type test. Increased
(12)
n = 154 bladder cancer patients, 60 benign
urological condition patients, 30 healthy
volunteers.
γ-synuclein,
catechol-omethyltransferase
and reticulin
EGF
n = 112 bladder cancer donors and n = 230
control donors.
Cytokeratinbased tests.
n = 54 bladder cancer donors (16 grade 1Ta,
38 grade 3 or T2> lesions) and n = 66
pathological or normal control samples
without bladder cancer.
n = 38 bladder cancer tissue sections, n = 23
normal urine samples and n = 19 bladder
cancer urine samples.
n=44 patients with bladder cancer as assessed
by cytoscopy and n=29 patients without
bladder cancer by cytoscopy.
n = 111 active bladder cancer patients, n = 76
disease-free controls.
Mucin-like
antigens (M344,
19A211 and
LDQ10 against
transitional cell
carcinoma in
exfoliated
urothelial cells )
and glycosylated
carcinoembryonic
antigen
Hyaluronic acid
Reference(s):
Utility‡
ELISA-based study:
Patients with detectable levels of urinary MMP1
had higher disease progression rates (P = 0.04).
ELISA and gelatin zymography, Detects early
tumour progression and elevated MMPs levels
correlate with tumour stage.
TIMP2
DEK-Protein
Study design and Diagnostic/Prognostic
n = 152 patients with either haematurias or
irritative voiding symptoms, n = 107 patients
under bladder cancer surveillance, n = 46 nonbladder cancer urinary tract pathologies, n =
20 healthy donors.
n = 150 donors post bladder cancer resection,
n = 114 donors with bladder-cancer symptoms
but not diagnosed.
n=30 bladder cancer patient urines (14 stage
(1)
(2)
(4)
(5)
(8)
(9)
(10)
and
hyaluronidase
Soluble Fas
receptor isoforms
Nuclear Matrix
Protein-22
(NMP22)
HSP60 and IL-13
Telomerase
pTa, 9 stage pT1, 5 stage pT2 and 2 with
carcinoma in situ transitional cell carcinoma)
and n=64 controls (55 with a history of
bladder cancer but no cytoscopic evidence of
bladder cancer at the time of sampling and 9
benign prostatic hyperplasia samples).
n = 188 patient urine samples including 31
non-cancer urologic controls and 10 healthy
controls.
hyaluronidase activity detects bladder cancer with
83.3% sensitivity and 78.1% specificity.
ELISA. ROC AUC for sFAS receptor = 0.757.
(13)
n = 43 newly diagnosed bladder cancer
patients, n = 58 recurrent bladder cancer, n =
77 remission patients and n = 25 healthy
controls.
Discovery set: n = 18 healthy donors, n = 20
haematuria patients, n = 50 non-muscleinvasive bladder cancer, n = 18 muscle
invasive bladder cancer. Validation set = n=19
control patients, n=21 bladder cancer.
n = 197 patients with urinary tract symptoms
and n=318 with diagnosis of bladder cancer.
ELISA based study (Bladder Chek). NMP22
detects bladder cancer with sensitivity of 88%
(newly diagnosed) and 57% (recurrent), mean
specificity = 67%.
ELISA based study. ROC AUC for IL-13 = 0.93
for prediction of bladder cancer. Combination of
HSP60 and IL-13 predicted bladder cancer with
positive predictive value = 74% and negative
predictive value = 76%.
Enzymatic Assay within urine. At 50 units cutoff for bladder cancer diagnosis, sensitivity =
87%, specificity = 70%.
ELISA-based study. UPK3A elevated in bladder
cancer. At cut-off of 0.0685 absorbance units,
sensitivity and specificity for UPK3A test = 83%.
ROC AUC = 0.907.
Immunofluorimetric assay. Increased MCM5protein predictive of bladder cancer. Minichromosome maintenance-5 protein test detects
bladder cancer with 69% specificity and 93%
negative predictive value. ROC AUC with
optimal MCM5 cut-off for diagnosis of bladder
cancer was 0.75.
ELISA-based study. Clusterin elevated in
bladder cancer and for detection of bladder cancer
sensitivity and specificity = 87.1% and 96.7%
respectively.
(14)
Uroplakin-3A
(UPK3A)
Urine samples, n = 32 healthy controls, n = 44
benign urological conditions, n = 122 bladder
cancer donors.
MiniChromosome
Maintenance-5
(MCM5) protein.
Total study group, n = 1677 consecutive
patients.
Clusterin
n = 68 bladder cancer donors, n = 61 benign
urological conditions.
(15)
(16)
(17)
(18)
(19)
†All samples are human and urine unless otherwise specified; n numbers refer to number with
condition unless otherwise stated,
‡Abbreviations used for proteomic techniques are defined within the text and list of common
abbreviations.
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