TERT promoter mutations are associated with distant metastases in upper tract urothelial carcinomas and serve as urinary biomarkers detected by a sensitive castPCR

Abstract

// Kun Wang 1, 2, * , Tiantian Liu 3, * , Nan Ge 4, * , Li Liu 5 , Xiaotian Yuan 2 , Jikai Liu 1 , Feng Kong 4 , Chang Wang 2 , Hongbo Ren 1 , Keqiang Yan 1 , Sanyuan Hu 6 , Zhonghua Xu 1 , Magnus Björkholm 2 , Yidong Fan 1 , Shengtian Zhao 4 , Cheng Liu 1 , Dawei Xu 2, 4 1 Department of Urology, Shandong University Qilu Hospital, Jinan, China 2 Department of Medicine, Division of Hematology and Centre for Molecular Medicine, Karolinska University Hospital Solna and Karolinska Institutet, Stockholm, Sweden 3 Department of Pathology, Shandong University School of Medicine, Jinan, China 4 Department of Urology and Central Research Laboratory, Shandong University Second Hospital, Jinan, China 5 Shandong University School of Nursing, Jinan, China 6 Department of General Surgery, Shandong University Qilu Hospital, Jinan, China * These authors contributed equally to this work Correspondence to: Shengtian Zhao, e-mail: zhaoshengtian@sdu.edu.cn Cheng Liu, e-mail: Cheng.Liu@qiluhospital.com Keywords: Cancer biomarkers, Promoter mutations, Telomerase, TERT, UTUC Received: September 02, 2014      Accepted: October 26, 2014      Published: December 09, 2014 ABSTRACT TERT promoter C228T and C250T mutations occur in various malignancies including bladder cancer (BC) and may serve as urinary tumor markers. However, the mutation association with clinical variables in upper tract urothelial carcinomas (UTUCs) is unclear. There is also a lack of sensitive tools to detect the minor mutant TERT promoter in bulk urinary DNA. Here we analyzed 220 UTUC patients [98 with renal pelvic carcinoma (RPC) and 122 with ureter carcinoma (UC)] and developed a Competitive Allele-Specific TaqMan PCR (castPCR) for urinary assay. We identified C228T or C250T mutations in 42 of 98 (43%) RPC and 23 of 122 (19%) UC tumors. Distant metastases were significantly correlated with UTUC patients harboring TERT promoter mutations ( P = 0.001). C228T were detected in 6/10 and 9/10 of urine samples from patients with mutation-carrying tumors using Sanger sequencing and castPCR, respectively. When urine samples from 70 BC patients were analyzed together, the sensitivity of urinary C228T assay was 89% and 50% for castPCR and Sanger sequencing, respectively ( P < 0.001). Collectively, TERT promoter mutations occur in UTUCs with a high frequency in RPCs and predict distant metastasis. castPCR assays of the mutation are a useful tool for urine-based diagnostics of urological malignancies.