Xiao Yang1, Yidong Cheng1, Qiang Lu1, Jifu Wei2, Haiwei Yang1, Min Gu1. 1. Department of Urology, The First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China. 2. Clinical Research Centre, The First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China.
Abstract
BACKGROUND: Circulating microRNAs are potential markers for disease detection. A novel class of small non-coding RNAs called Piwi-interacting RNAs (piRNAs) has been recently reported to participate in the epigenetic regulation of cancers and other diseases. This study aims to discover blood-based piRNAs which can be used as markers for disease detection and monitoring. MATERIALS AND METHODS: We selected five piRNAs for detection, namely, has-piR-651, has-piR-823, has-piR-36707, has-piR-36741 and has-piR-57125. Serum or plasma samples were used to isolate small RNAs, including the piRNAs. The extracted small RNAs were reverse-transcribed in the presence of a poly-A polymerase with an oligo-dT adaptor, and quantitative real-time PCR (qRT-PCR) was applied to measure the levels of piRNAs. Room-temperature incubation and repetitive freeze-thaw cycles were performed to measure the stability of the piRNAs. RESULTS: Unlike the four other piRNAs, has-piR-57125 was present in both the serum and plasma samples. Regardless of the serum or plasma samples, qRT-PCR analysis indicated that the Ct values showed no remarkable variation with prolonged incubation time (P > 0.05). We also detected the Ct values of the samples with repetitive freeze-thaw cycles and observed a similar trend (P > 0.05) among the samples with diverse freeze-thaw cycles. CONCLUSION: This study is the first to report that piRNAs are stably expressed in human serum or plasma samples. Therefore, piRNAs can serve as valuable blood-based biomarkers for disease detection and monitoring.
BACKGROUND: Circulating microRNAs are potential markers for disease detection. A novel class of small non-coding RNAs called Piwi-interacting RNAs (piRNAs) has been recently reported to participate in the epigenetic regulation of cancers and other diseases. This study aims to discover blood-based piRNAs which can be used as markers for disease detection and monitoring. MATERIALS AND METHODS: We selected five piRNAs for detection, namely, has-piR-651, has-piR-823, has-piR-36707, has-piR-36741 and has-piR-57125. Serum or plasma samples were used to isolate small RNAs, including the piRNAs. The extracted small RNAs were reverse-transcribed in the presence of a poly-A polymerase with an oligo-dT adaptor, and quantitative real-time PCR (qRT-PCR) was applied to measure the levels of piRNAs. Room-temperature incubation and repetitive freeze-thaw cycles were performed to measure the stability of the piRNAs. RESULTS: Unlike the four other piRNAs, has-piR-57125 was present in both the serum and plasma samples. Regardless of the serum or plasma samples, qRT-PCR analysis indicated that the Ct values showed no remarkable variation with prolonged incubation time (P > 0.05). We also detected the Ct values of the samples with repetitive freeze-thaw cycles and observed a similar trend (P > 0.05) among the samples with diverse freeze-thaw cycles. CONCLUSION: This study is the first to report that piRNAs are stably expressed in human serum or plasma samples. Therefore, piRNAs can serve as valuable blood-based biomarkers for disease detection and monitoring.
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