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Published on: November 28, 2018
Detection of stably expressed piRNAs in human blood
Xiao Yang1, Yidong Cheng1, Qiang Lu1
1Department of Urology, The First Affiliated Hospital of Nanjing Medical University Nanjing 210029, China.
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.
Insights
Piwi-interacting RNAs (piRNAs) are stable in blood samples, making them promising biomarkers for disease detection and monitoring. This study identified specific piRNAs that remain consistent under various conditions, supporting their diagnostic potential.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- Circulating microRNAs are emerging as critical biomarkers for disease detection.
- Piwi-interacting RNAs (piRNAs), a class of small non-coding RNAs, are implicated in the epigenetic regulation of diseases.
- This research focuses on identifying blood-based piRNAs for diagnostic and monitoring applications.
Purpose of the Study:
- To discover and validate blood-based piRNAs as potential biomarkers for disease detection and monitoring.
- To investigate the stability of specific piRNAs in human serum and plasma samples.
Main Methods:
- Selected five piRNAs (has-piR-651, has-piR-823, has-piR-36707, has-piR-36741, has-piR-57125) for analysis.
- Isolated small RNAs from serum and plasma, followed by reverse transcription and quantitative real-time PCR (qRT-PCR).
- Assessed piRNA stability through room-temperature incubation and repetitive freeze-thaw cycles.
Main Results:
- Has-piR-57125 was detected in both serum and plasma samples.
- Quantitative real-time PCR showed no significant variation in Ct values for piRNAs with prolonged incubation or repetitive freeze-thaw cycles.
- PiRNA levels remained stable across different storage and handling conditions.
Conclusions:
- This study provides the first evidence of stable piRNA expression in human serum and plasma.
- PiRNAs demonstrate potential as robust blood-based biomarkers for disease detection and monitoring.
- The stability of piRNAs supports their utility in clinical diagnostics.
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