High-Throughput Sequencing Strategy for miR-146b-regulated circRNA Expression in Hepatic Stellate Cells.
High-Throughput Sequencing Strategy for miR-146b-regulated circRNA Expression in Hepatic Stellate Cells.
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DOI：
10.12659/msm.910807
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发表时间：
2018-12-01
期刊：
影响因子：
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通讯作者：
Xiang T
中科院分区：
文献类型：
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作者：
Cheng N;Xiao J;Ge S;Li J;Huang J;Wu X;Zhang S;Xiang T
本研究旨在检测和分析miR-146b介导的环状RNA在肝星状细胞中的表达。实验分为对照组和siRNA-miR-146b组。用实时定量逆转录聚合酶链式反应(qRT-PCR)证实siRNA-miR-146b的干扰效率，收集细胞，收集总RNA进行高通量测序。预测了miRNA靶向的身体。最后，通过qRT-PCR验证了5个CircRNAs的表达。SiRNA-miR-146b组的miR-146b表达明显低于对照组。原始测序数据和处理后数据的质量均满足分析要求，miR-146b的缺失对CircRNAs的表达有调节作用。与对照组相比，miR-146b组有18个CircRNA表达上调，77个CircRNAs表达下调。基因预测表明，hsa_Circ1887是miRNA和CircRNA调控网络中最大的接触点。QRT-PCR结果显示，与对照组相比，siRNA-miR-146b组的RNO-CircRNA-469、RNO-CircRNA-1138、RNO-CircRNA-2168和RNO-CircRAN-1907显著降低，而CircRNA-1984显著上调，这与高通量测序技术的结果一致。MIR-146b可调节肝干细胞CircRNAs的表达，可能参与肝纤维化的形成和发展。
This study was designed to detect and analyze miR-146b-mediated circular RNA (circRNA) expression in hepatic stellate cells. The experiment was divided into a control group and a siRNA-miR-146b group. The interference efficiency of siRNA-miR-146b was confirmed by real-time quantitative reverse transcription PCR (qRT-PCR) and the cells were collected, and total RNA was collected for high flux sequencing. The miRNA-targeted carcass were predicted. Finally, the expression of 5 circRNAs was verified by qRT-PCR. miR-146b expression in the siRNA-miR-146b group was significantly lower than that in the control group. The quality of the original sequencing data and the processed data satisfied with the analysis, and the expression of circRNAs was modulated after the reduction of miR-146b. Among them, 18 circRNAs were upregulated, while 77 circRNAs were downregulated in the miR-146b group compared with the control group. The gene prediction showed that hsa_circ1887 was the largest contact point in miRNA and circRNA regulatory networks. qRT-PCR showed that rno-circRNA-469, rno-circRNA-1138, rno-circRNA-2168 and rno-circRAN-1907 were significantly reduced, while circRNA-1984 was significantly promoted in the siRNA-miR-146b group compared with the control group, which were consistent with the measurements by high-throughput sequencing technique. miR-146b could regulate the expression of circRNAs in HSCs, which might take part in the formation and development of hepatic fibrosis.