Article

Comparison of cytokeratin 20 RNA and angiogenin in voided urine samples as diagnostic tools for bladder carcinoma.

Oncology Diagnostic Unit, Biochemistry Department, Faculty of Medicine, Ain Shams University, Abbassia, Cairo, Egypt.
Clinical Biochemistry (Impact Factor: 2.28). 10/2004; 37(9):803-10. DOI: 10.1016/j.clinbiochem.2004.05.027
Source: PubMed

ABSTRACT We evaluated the diagnostic efficacy of urinary angiogenin (ANG) and cytokeratin 20 (CK-20) mRNA in comparison with voided urine cytology in the detection of bladder cancer patients.
A total of 97 Egyptian patients provided a single voided urine sample for ANG, CK-20 and cytology before cystoscopy. Of the 97 cases, 63 were histologically diagnosed as bladder cancer; 33 with transitional cell carcinoma (TCC) and 30 with squamous cell carcinoma (SCC), whereas the remaining 34 had benign urological disorders. A group of 46 healthy volunteers were also included in this study. Voided urine was centrifuged and the supernatant was used for estimation of ANG by EIA and confirmed by Western blotting (WB). The urine sediment was used for cytology and RNA extraction. CK-20 RNA was detected by RT-PCR.
The best cutoff value for ANG was calculated by a ROC curve as 322.7 ng/mg protein. The median urinary ANG level in bladder carcinoma, benign urological disorders and healthy volunteer groups was: 802.7, 425 and 33 pg/mg protein, respectively. The positivity rate for urinary CK-20 mRNA of the control, benign and malignant groups was 0%, 2.9% and 82.3%, respectively (P = 0.000); while the rates for ANG were 11.6%, 54.8% and 75.4%, respectively (P = 0.000). There was no significant difference in positivity rates of CK-20 and ANG with respect to sex, smoking, schistosomiasis, urine cytology, tumor grade, tumor stage, hematuria or pus cells. The overall sensitivity and specificity were 71.4% and 90% for voided urine cytology, 75.4% and 70.3% for ANG, and 82.3% and 98.8% for CK-20. Combined sensitivity of voided urine cytology with ANG and CK-20 together (98.2%) was higher than either the combined sensitivity of voided urine cytology with ANG (96.5%) or with CK-20 (91.6%) or than that of the biomarker alone. We demonstrated significant positive correlation between CK-20 positivity with age (P = 0.043) and nodal involvement (P = 0.037); however, there was no significant correlation between CK-20 and ANG with the other clinicopathological parameters.
Our data indicate that CK-20 and ANG in voided urine had higher sensitivities compared to voided urine cytology. However, when specificity was considered, CK-20 alone had superior sensitivity and specificity compared to ANG and voided urine cytology.

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    • "The pathogenetic profile of urinary bladder cancer has led to substantial interest in the development of diagnostic and prognostic indicator or tumor markers of cancer initiation and progression in both schistosome and non schistosome bladder cancer (Saied et al., 2006; El-Salahy, 2002; Khaled et al., 2001; El-Ahmady, 1999; Ahmed, 1995; El-Sewedy, 1978; Eissa, 2008, 2004). Differential expression of these tumor markers in relation of schistosome associated bladder cancer when compared to non-schistosome urinary bladder cancer also lends credence to the etiological link between the parasite and urinary bladder cancer. "
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    • "Special interest has been generated in non-invasive objective urinary markers, which are helpful in the diagnosis of bladder cancer. Unfortunately, because a single tumor marker with sensitivity and negative predictive value of 100% that could replace cystoscopy does not exist, the role of urinary markers is still adjunctive to cystoscopy, substituting or eliminating the need for cytology in certain indications [13] [19] [20] [30] [31]. As a result, urologist attempted to choose different urinary markers as a combined assay to increase the sensitivity for the detection of bladder cancer. "
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    ABSTRACT: We evaluated the diagnostic efficacy of urinary CD44 and cytokeratin 20 (CK20) mRNA in comparison with voided urine cytology (VUC) for the detection of bladder cancer. A total of 136 Egyptian patients provided a single voided urine sample for CD44, CK20 mRNA and VUC before cystoscopy. Of the 136 cases, 111 were histologically diagnosed as bladder cancer whereas the remaining 25 had benign urological disorders. A group of 20 healthy volunteers was also included in this study. Voided urine was centrifuged and the urine sediment was used for cytology, estimation of CD44 by ELISA and RNA extraction. CK20 mRNA was detected by conventional RT-PCR and quantitative real-time RT-PCR. The best cutoff values for CD44 and relative CK20 mRNA detected by real-time RT-PCR were calculated by receiver operating characteristic curve. The positivity rates and the mean ranks for CD44 and CK20 mRNA showed significant difference among the three investigated groups (p=0.001). Quantitative real-time RT-PCR results were comparable to conventional RT-PCR for the detection of CK20 mRNA. The positivity rate of CD44 was significantly associated with schistosomiasis and urine cytology. The overall sensitivity and specificity were 52.3% and 88.9% for VUC, 63.1% and 88.9% for CD44, and 82.0% and 97.8% for CK20 mRNA. Combined sensitivity of VUC with CD44 and CK20 mRNA together (95.5%) was higher than either the combined sensitivity of VUC with CD44 (78.4%) or with CK20 mRNA (91.0%) or than that of the biomarker alone. Urinary CD44 and CK20 mRNA had higher sensitivities compared to VUC. However, when the diagnostic efficacy was considered, CK20 mRNA by either conventional RT-PCR or real-time RT-PCR had the highest sensitivity and specificity compared to CD44 and VUC.
    Clinical biochemistry 10/2008; 41(16-17):1335-41. DOI:10.1016/j.clinbiochem.2008.08.085 · 2.28 Impact Factor
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    • "Western blot was performed according to Sambrook (Sambrook et al., 1998) and adapted by Eissa (Eissa et al., 2004). Briefly, proteins from 60 µg cell lysates were separated by 12% sodium dodecyl sulfatepolyacrylamide gel electrophoresis. "
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