Development of 3-Hydroxyanthranilic acid-based Integrated Non-invasive Biosensor for Bladder Cancer Detection
Yuh-Shyan Tsai1, Yeong-Chin Jou2, Yen-Ping Tsai1, Bin-Da Liu3, Hung-In Lin4, Chia-Lin Wei3, Syue-Yi Chen5, Hsin-Tzu Tsai1, Chien-Hui Ou1, Wen-Horng Yang1, Tzong-Shin Tzai1
Department of Urology1, Department of Electric Engineering3, College of Medicine and Hospital, National Cheng Kung University, Tainan, Taiwan
Department of Urology2, and Medical Researh5, Chia-Yi Christian Hospital, Chia-Yi, Taiwan
, National Cheng Kung University, Tainan, Taiwan
Department of Chemical and Material Engineering4, National Kaohsiung University, Kaohsiung, Taiwan
Introduction and Objectives: Bladder cancer is a common human malignancy and exhibits a life-long risk of disease recurrence and progression. It is continuing to search some simple, innovative biomarker to monitor the disease status in order to diminish the suffering during cystoscopic follow-up. The metabolite of tryptophan after indoleamine 2,3-dioxygenase (IDO) digestion, 3-Hydroxyanthranilic acid (3-HAA) is conventionally thought to be a potential biomarker for bladder cancer occurrence. The aim to study is to investigate the diagnostic potential of an integrated a 3-HAA-based biosensor for urothelial carcinoma of the upper tract and urinary bladder.
Materials and Methods: Human urothelial cancer cell lines and human urothelial carcinoma tissues as well as adjacent benign tissues were available for exploring IDO expression, including western blotting and immunohistochemical staining. Patients who received urological surgery were enrolled for urine 3-HAA testing using an integrating biosensor for 3-HAA. Some of urine specimens were investigated with high performance liquid chromatography (HPLC) assay.
Results: From western blotting assays, eight human urothelial carcinoma cell lines exhibited more IDO expression than the immortalized cell SV-HUC. Both of urothelial carcinoma of urinary bladder and upper urinary tract exhibited more IDO immunoreactivity than those of the adjacent benign bladder, ureteral or cortical tissues (chi-square test, p=0.0073). There is a moderate correlation for urine 3-HAA measurement based on between HPLC and the biosensor assays (r2=0.47, p=0.0033). Besides, the 3-HAA content within the cultured media of TCCSUP and BFTC905 measured with biosensors significantly increased with incubation time (p <0.0001). Finally, Patients with urothelial carcinoma of bladder and upper tract have higher urine 3-HAA levels than those without recurrence or benign urological disease, such as BPH, or hernia (unpaired t-test, p=0.028), except for urolithiasis.
Conclusion: The integrated biosensor exhibited a modest accuracy in urine 3-HAA detection. Both of urothelial carcinoma of urinary bladder and upper tract exhibited higher IDO expression and its metabolite 3-HAA in urine.