Transcriptome analyses of urine RNA reveal tumor markers for human bladder cancer: validated amplicons for RT-qPCR-based detection
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Josephine Dubois1, Jacqueline Rueger1, Bernhard Haubold2, Rosel Kretschmer-Kazemi Far1 and Georg Sczakiel1
1 Institut für Molekulare Medizin, Universität zu Lübeck and UKSH, Campus Lübeck, Lübeck D-23538, Germany
2 Max-Planck-Institute for Evolutionary Biology, Department of Evolutionary Genetics, Ploen 24306, Germany
Keywords: tumor diagnostics; non-invasive; urine RNA; biomarker; transcriptome analysis
Received: March 17, 2021 Accepted: April 19, 2021 Published: May 11, 2021
Non-invasive clinical diagnostics of bladder cancer is feasible via a set of chemically distinct molecules including macromolecular tumor markers such as polypeptides and nucleic acids. In terms of tumor-related aberrant gene expression, RNA transcripts are the primary indicator of tumor-specific gene expression as for polypeptides and their metabolic products occur subsequently. Thus, in case of bladder cancer, urine RNA represents an early potentially useful diagnostic marker.
Here we describe a systematic deep transcriptome analysis of representative pools of urine RNA collected from healthy donors versus bladder cancer patients according to established SOPs. This analysis revealed RNA marker candidates reflecting coding sequences, non-coding sequences, and circular RNAs. Next, we designed and validated PCR amplicons for a set of novel marker candidates and tested them in human bladder cancer cell lines. We identified linear and circular transcripts of the S100 Calcium Binding Protein 6 (S100A6) and translocation associated membrane protein 1 (TRAM1) as highly promising potential tumor markers.
This work strongly suggests exploiting urine RNAs as diagnostic markers of bladder cancer and it suggests specific novel markers. Further, this study describes an entry into the tumor-biology of bladder cancer and the development of gene-targeted therapeutic drugs.
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