Novel combination markers for predicting progression of nonmuscle invasive bladder cancer Yun-Sok Ha 1 , Ji Sang Kim 1 , Hyung-Yoon Yoon 1 , Pildu Jeong 1 , Tae-Hwan Kim 2 , Seok-Joong Yun 1 , Sang-Cheol Lee 1 , Gi-Young Kim 3 , Yung-Hyun Choi 4 , Sung-Kwon Moon 5 , Isaac Yi Kim 6 and Wun-Jae Kim 1 1 Department of Urology, College of Medicine, Chungbuk National University, Cheongju, South Korea 2 Department of Urology, College of Medicine, Kyungpook National University, Daegu, South Korea 3 Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju, South Korea 4 Department of Biochemistry, Dongeui University College of Oriental Medicine, Busan, South Korea 5 Department of Food and Biotechnology, Chungju National University, Chungju, Chungbuk, South Korea 6 Dean and Betty Gallo Prostate Cancer Center and Section of Urologic Oncology, Division of Urology, Department of Surgery, The Cancer Institute of New Jersey, Robert Wood Johnson Medical School, New Brunswick, NJ To identify prognostic markers in nonmuscle invasive bladder cancer (NMIBC), the combined effect of RUNX3 and MGC17624 for predicting NMIBC progression was assessed. RUNX3 promoter methylation was examined using methylation specific- polymerase chain reaction (MS-PCR). MGC17624 mRNA expression was evaluated by real-time PCR. Patients were divided into three groups according to the status of the two genes and the prognostic effects of these markers were evaluated. The median follow-up period was 57.8 months (range, 9.1–189.7). The mRNA expression level of MGC17624 was significantly lower in patients with positive RUNX3 methylation than in those with negative methylation (p 5 0.047). Kaplan–Meier estimates showed significant differences in time-to-progression between the genetic combination predictors (log-rank test; p < 0.001). Patients with a poor predictive combination were at a significantly higher risk for progression [Hazard ratio (HR), 22.579] than patients with a good predictive combination in multivariate Cox regression analysis. In the subgroup analysis, a poor predictive combination accurately estimated progression in patients with intravesical therapy (HR, 20.081) and in those who experienced recurrence (HR, 54.233). Assessment of the status of RUNX3 and MGC17624 in combination was identified as a reliable method for predicting NMIBC progression. Bladder cancer has diverse biological and functional charac- teristics, which are determined by the multistep accumulation of genetic and epigenetic alterations through multiple genetic disorders. 1 These molecular alterations result in uncontrolled cell proliferation, decreased cell death, invasion and metasta- sis, and may therefore influence the patient’s prognosis. Con- ventional clinicopathological parameters such as grade and stage are widely used to predict the clinical outcome in blad- der cancer, but their predictive ability is limited, especially with respect to prospective risk assessment in individual patients. 2,3 The need to establish which bladder cancer will recur, progress or metastasize has led to the identification of a variety of potential prognostic markers for bladder cancer patients. However, none of the biomarkers reported to date have shown sufficient sensitivity and specificity for detecting the whole spectrum of bladder cancer diseases in routine clinical practice. 4,5 Intensive efforts to find an accurate prognostic factor for bladder cancer led to the identification of RUNX3. RUNX3 is a tumor suppressor gene and its inactivation by DNA hyper- methylation has been reported in various tumors. 6,7 In previ- ous studies, promoter hypermethylation of RUNX3 was found to be closely associated with tumor grade, invasiveness and prognosis of bladder cancer. 8,9 Validation of the initial results with long-term follow-up of additional cases confirmed the reliability of the study results and suggested that RUNX3 could be a valuable gene as a prognostic marker for bladder cancer. Furthermore, the use of RUNX3 as a target for epige- netic therapy will offer the possibility of reversing epigenetic changes and restoring gene function. 10 Multiple genetic alterations are involved in the transfor- mation of a normal cell into a malignant and finally meta- static phenotype. The assessment of multiple markers is therefore more accurate to describe the biological phenotype of a particular cancer. To overcome the limitations of single Key words: nonmuscle invasive bladder cancer, prognosis, MGC17624, RUNX3 Grant sponsor: National Research Foundation of Korea (NRF); Grant number: 2011-0001042; Grant sponsor: Ruth Estrin Goldberg Memorial Foundation, the Tanzman Foundation, the Jon Runyan Score for the Cure DOI: 10.1002/ijc.27319 History: Received 3 May 2011; Accepted 14 Oct 2011; Online 25 Oct 2011 Correspondence to: Wun-Jae Kim, Department of Urology, College of Medicine, Chungbuk National University, 62 Kaesin-dong, Heungduk-gu, Cheongju 361-763, South Korea, Tel.: þ82-43-269-6137, Fax: þ82-43-269-6144, E-mail: wjkim@chungbuk.ac.kr Early Detection and Diagnosis Int. J. Cancer: 000, 000–000 (2011) V C 2011 UICC International Journal of Cancer IJC