Yonsei Med J.  2016 Jul;57(4):865-871. 10.3349/ymj.2016.57.4.865.

Metabolic Pathway Signatures Associated with Urinary Metabolite Biomarkers Differentiate Bladder Cancer Patients from Healthy Controls

• Kim WT
• Yun SJ
• Yan C
• Jeong P
• Kim YH
• Lee IS
• Kang HW
• Park S
• Moon SK
• Choi YH
• Choi YD
• Kim IY
• Kim J
• Kim WJ

• KMID: 2374116
• DOI: http://doi.org/10.3349/ymj.2016.57.4.865

Abstract

PURPOSE
Our previous high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry study identified bladder cancer (BCA)-specific urine metabolites, including carnitine, acylcarnitines, and melatonin. The objective of the current study was to determine which metabolic pathways are perturbed in BCA, based on our previously identified urinary metabolome.
MATERIALS AND METHODS
A total of 135 primary BCA samples and 26 control tissue samples from healthy volunteers were analyzed. The association between specific urinary metabolites and their related encoding genes was analyzed.
RESULTS
Significant alterations in the carnitine-acylcarnitine and tryptophan metabolic pathways were detected in urine specimens from BCA patients compared to those of healthy controls. The expression of eight genes involved in the carnitine-acylcarnitine metabolic pathway (CPT1A, CPT1B, CPT1C, CPT2, SLC25A20, and CRAT) or tryptophan metabolism (TPH1 and IDO1) was assessed by RT-PCR in our BCA cohort (n=135). CPT1B, CPT1C, SLC25A20, CRAT, TPH1, and IOD1 were significantly downregulated in tumor tissues compared to normal bladder tissues (p<0.05 all) of patients with non-muscle invasive BCA, whereas CPT1B, CPT1C, CRAT, and TPH1 were downregulated in those with muscle invasive BCA (p<0.05), with no changes in IDO1 expression.
CONCLUSION
Alterations in the expression of genes associated with the carnitine-acylcarnitine and tryptophan metabolic pathways, which were the most perturbed pathways in BCA, were determined.

Keyword

Bladder cancer; urine metabolites; gene expression; qRT-PCR; diagnostic marker

MeSH Terms

Aged
Biomarkers/metabolism
Carcinoma, Transitional Cell/genetics/*metabolism/pathology
Carnitine/*analogs & derivatives/genetics/metabolism
Case-Control Studies
Female
Humans
Male
Metabolic Networks and Pathways/*physiology
Middle Aged
RNA, Messenger/metabolism
Real-Time Polymerase Chain Reaction
Urinary Bladder Neoplasms/genetics/*metabolism/pathology
Biomarkers
Carnitine
RNA, Messenger

Figure

• 

  Fig. 1 Jitter plots showing mRNA levels in normal controls and patients with NMIBC or MIBC. RT-PCR analysis of the mRNA expression of CPT1A, CPT1B, CPT1C, CPT2, SLC25A20, CRAT, TPH1, and IDO1 was performed as described in the Materials and Methods section. p values were calculated using the Mann-Whitney U test. NMIBC, non-muscle invasive bladder cancer; MIBC, muscle invasive bladder cancer; IDO, indoleamine 2,3-dioxygenase; CPT, carnitine palmitoyltransferase; CRAT, carnitine O-acetyltransferase; TPH, tryptophan hydroxylase; SLC25A20, solute carrier family 25 [carnitine/acylcarnitine translocase (CACT)], member 20.

• 

  Fig. 2 Schematic pathway of bladder cancer-associated metabolic signatures in bladder cancer patients. CPT, carnitine palmitoyltransferase; CACT, carnitine/acylcarnitine translocase; CRAT, carnitine O-acetyltransferase; TPH, tryptophan hydroxylase; IDO, indoleamine 2,3-dioxygenase.

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