Determination of Serum Exosomal H19 as a Noninvasive Biomarker for Bladder Cancer Diagnosis and Prognosis

BACKGROUND: Long noncoding RNAs (lncRNA) contained in exosomes have been recognized as promising stable biomarkers in cancers. H19 is a well-known oncogenic lncRNA, but whether extracellular H19 is contained in exosomes and the clinical significance of exosomal H19 are poorly understood. The aim of this study was to assess serum-derived exosomal H19 lncRNA as a cancer predictive marker. MATERIAL/METHODS: Exosomes from serum of bladder cancer (BC) patients were isolated using the ExoQuick purification method and identified using transmission electron microscopy and nanoparticle tracking analysis. RT-qPCR was used for the measurement of H19 expression in serum or tissue samples. RESULTS: Serum H19 was little influenced by the treatment of RNase A alone but was dramatically downregulated when treated with RNase A and Triton ×100 simultaneously. The concentration of H19 was significantly higher in serum exosomes than in exosome-depleted supernatants in serum. Then, we validated that exosomal H19 is stable in serum by exposing serum samples to prolonged conditions of room temperature, 4°C, multiple freeze-thaw cycles, and low/high pH. Serum exosomal H19 of BC patients was positively correlated with total H19 level in paired BC tissues, and exosomal H19 was significantly downregulated in postoperative samples when compared to the paired preoperative samples. In addition, exosomal H19 level was significantly increased in serum of BC patients when compared to healthy people and benign disease patients. More importantly, Kaplan-Meier survival curve analysis showed that higher serum exosomal H19 level in BC patients was correlated with poorer survival. CONCLUSIONS: Detection of serum exosomal H19 shed light on using exosomal lncRNAs as a noninvasive diagnostic and prognostic biomarker for BC patients.