Cox-2 inhibition demonstrates potent anti-proliferative effects on bladder cancer in vitro

Abstract

Purpose: Determine the in vitro effect of Rofecoxib in vitro in regards to cell growth, apoptotic activity, and prostaglandin production. Introduction and Objective: Rofecoxib (VIOXX) is a non-steroidal anti-inflammatory agent that selectively inhibits cyclooxygenase-2 (COX-2). COX-2 is a critical enzyme in the conversion of arachidonic acid to prostaglandin E2. The inducible isoform of COX-2 is overexpressed in many gastrointestinal and genitourinary tract tumors. We hypothesized that in vitro treatment with rofecoxib would inhibit the growth of bladder cancer cells by apoptotic pathways. Methods: Two human bladder cancer cell lines (T24, mutant p53, and TCCSUP, p53 wild type) were grown in culture using standard techniques and treated with rofecoxib at a dose of 125 μg/well. Cell viability was measured by MTT at 24 and 72 hours. Apoptosis was evaluated using a cell death detection kit. Prostaglandin E2 (PGE2) was measured by ELISA. Statistical analysis was performed by ANOVA. Results:TABLE—ABSTRACT P11CellsApoptosis% Cell inhibitionPGE2 (pg/ml)T24 CtlN.A.N.A.147.4 ± 4.3T24 24 hrs226.7 ± 57.0%∗18.1 ± 10.4%84.1 ± 3.8∗T24 CtlN.A.N.A.234.5 ± 5.6T24 72 hrsN.A.64.4 ± 3.2%∗87.9 ± 6.7∗TCC CtlN.A.N.A.4669.9 ± 27.4TCC 24 hrs110.4 ± 28.4%3.7 ± 7.5%4218.3 ± 24.8∗TCC CtlN.A.N.A.4622.5 ± 47.1TCC 72 hrsN.A.67.9 ± 2.9%∗3622.1 ± 42.6∗∗P < 0.01 compared to untreated controls.Conclusions: Selective COX-2 inhibition, using the well-tolerated and commercially available rofecoxib (VIOXX), reduced the growth of human bladder cancer in vitro by apoptotic mechanisms. Further in vivo and human studies are warranted to evaluate the safety and clinical utility of this agent in patients with bladder cancer.