The Real-Time Dynamic Monitoring of microRNA Function in Cholangiocarcinoma

BACKGROUND Although many studies have confirmed a relationship between microRNAs (miRNAs) and cholangiocarcinoma (CCA), the real-time dynamics of miRNA function have not been examined. METHODS miRNA reporter constructs were generated using a recombinant adeno-associated virus vector, which contained complementary sequences for six miRNAs (miR-200a, miR-200b, miR-21, miR-146a, miR-155, and miR-221), along with two independent expression cassettes encoding the fluorescent reporter genes Fluc and Gluc. The spatio-temporal function of each miRNA was monitored both in CCA and control tissues. RESULTS All miRNAs participated in CCA development, with distinct patterns of expression over time. The activity of miR-21 was significantly lower in female T3N0M0 CCA tissue relative to controls at three time points, yet was higher in two male T3N1M0 CCA tissues. The difference in miR-200b function between two male T3N1M0 CCA tissues and their corresponding controls peaked at 24 h, while function in a female T3N0M0 CCA was detected only at 72 h. The four remaining miRNAs (miR-200a, miR146a, miR-155, and miR-221) displayed patient-specific activity patterns in both CCA and control tissues. CONCLUSION Significant variability was observed in the temporal function of all six miRNAs, which may play an important role in the development of CCA.