Detection and molecular staging of bladder cancer using real-time RT-PCR for gelatinases (MMP-2, MMP-9) and TIMP-2 in peripheral blood

• Published in: Actas urologicas españolas 2011, Vol.35 (3), p.127-136
• Main Authors: Angulo, J.C, Ferruelo, A, Rodríguez-Barbero, J.M, Núñez, C, de Fata, F.R, González, J
• Format: Article
• Language: eng ; spa
• Subjects: Adult; Aged; Aged, 80 and over; Bladder neoplasia; Estadificación molecular; Female; Humans; Male; Matrix Metalloproteinase 2 - blood; Matrix Metalloproteinase 9 - blood; Middle Aged; MMP-2; MMP-9; Molecular staging; Neoplasia vesical; Neoplasm Staging; Reverse Transcriptase Polymerase Chain Reaction - methods; RT-PCR; TIMP-2; Tissue Inhibitor of Metalloproteinase-2 - blood; Urinary Bladder Neoplasms - blood; Urinary Bladder Neoplasms - pathology; Urology
• ISSN: 2173-5786; 2173-5786; 1699-7980
• DOI: 10.1016/S2173-5786(11)70035-X

Abstract Introduction Molecular staging of bladder cancer based on the detection of mRNA of urothelial specific genes in circulating cancer cells has been inconclusive. We analyze whether real-time RT-PCR evaluation of gelatinases (MMP-9, MMP-2) and TIMP-2 in peripheral blood allows diagnosing and characterizing patients with bladder neoplasm. Materials and method Total RNA is extracted from circulating blood cells in 42 individuals (11 healthy controls, 31 patients with bladder cancer in different stages) and real-time RT-PCR performed using specific primers for MMP-9, MMP-2, TIMP-2 and ribosomal 18S. The quantification values of mRNA are described as relative to 18S mRNA (ΔΔCt method) and the results are blindly compared with data obtained from histological diagnosis and clinical staging. Results Normalized levels of MMP-9 and MMP-2 mRNA are higher in patients with cancer than controls (1.82 ± 0.6 times and 2.7 ± 0.6 times, respectively; P < 0.05). Patients with metastatic disease also have increased MMP-9, MMP-2 and TIMP-2 mRNA levels (9.6 ± 0.20 times, 5.22 ± 0.26 times and 1,97 ± 0.22 times, respectively; P < 0.05). MMP-9 and MMP-2 are also associated with advanced clinical stage and grade (P < 0.05). A ratio between variables that increases the ability to segregate patients with Ta, T1, T2-4M0 and T2-4M1 tumors is proposed. Conclusions Both non-invasive bladder tumor recognition and molecular staging of the disease is possible using real-time RT-PCR-based detection of gelatinases and TIMP-2 in peripheral blood. The ability to distinguish metastatic disease is higher for MMP-9 but MMP-2 discriminates better levels of tumor invasion. Further investigation in this field could yield promising results regarding molecular evaluation of bladder neoplasia.