Detection of circulating tumor cells in bladder cancer using multiplex PCR assays

• Published in: Anticancer research Vol. 34; no. 12; pp. 7415 - 7424
• Main Authors: Leotsakos, Ioannis, Dimopoulos, Panagiotis, Gkioka, Eliona, Msaouel, Pavlos, Nezos, Adrianos, Stravodimos, Konstantinos G, Koutsilieris, Michael, Constantinides, Constantinos A
• Format: Journal Article
• Language: English
• Published: Greece 01.12.2014
• Subjects: Aged; Biomarkers, Tumor - blood; Biomarkers, Tumor - genetics; Biomarkers, Tumor - urine; Disease-Free Survival; Female; Humans; Inhibitor of Apoptosis Proteins - genetics; Inhibitor of Apoptosis Proteins - urine; Keratin-19 - blood; Keratin-20 - blood; Keratin-20 - genetics; Keratin-20 - urine; Male; Multiplex Polymerase Chain Reaction - methods; Neoplasm Grading; Neoplasm Staging - methods; Neoplastic Cells, Circulating - pathology; Receptor, Epidermal Growth Factor - blood; RNA, Messenger - urine; Telomerase - genetics; Telomerase - urine; Urinary Bladder Neoplasms - blood; Urinary Bladder Neoplasms - pathology; Urinary Bladder Neoplasms - urine; h-TERT; SURVIVIN; bladder cancer; CK20; Circulating tumor cells; EGFR; PCR; CK19

The aim of this study was to develop multiplex-PCR assays for the detection of circulating tumor cells in peripheral blood and urine samples of patients with bladder cancer. Peripheral blood and urine samples were collected from 208 patients (169 patients and 39 healthy volunteers). After RNA extraction and cDNA synthesis, the samples were analyzed for the expression of cytokeratin 19 (CK19), CK20 and epidermal growth factor receptor (EGFR) mRNA in blood and for SURVIVIN, human telomerase reverse transcriptase (hTERT), cytokeratin 20 (CK20) mRNA in urine, using multiplex-PCR assays. EGFR and CK20 alone or in combination as well as all urine markers correlated well with histological grade. hTERT correlated well with primary tumor size T≥3. Patients with positive urine markers had significantly worse progression-free survival. Multiplex-PCR assays can be a useful tool for staging and monitoring purposes in patients with bladder cancer.