Department of Infectious Diseases, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China (mainland).
Department of Ultrasound, Jiangxi Maternal and Child Health Hospital, Nanchang, Jiangxi, China (mainland).
Med Sci Monit. 2018 Dec 1;24:8699-8706. doi: 10.12659/MSM.910807.
BACKGROUND This study was designed to detect and analyze miR-146b-mediated circular RNA (circRNA) expression in hepatic stellate cells. MATERIAL AND METHODS 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. RESULTS 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. CONCLUSIONS miR-146b could regulate the expression of circRNAs in HSCs, which might take part in the formation and development of hepatic fibrosis.
本研究旨在检测和分析肝星状细胞中 miR-146b 介导的环状 RNA（circRNA）表达。
实验分为对照组和 siRNA-miR-146b 组。通过实时定量逆转录 PCR（qRT-PCR）确认 siRNA-miR-146b 的干扰效率，并收集细胞，提取总 RNA 进行高通量测序。预测 miRNA 的靶基因。最后，通过 qRT-PCR 验证 5 个 circRNA 的表达。
siRNA-miR-146b 组 miR-146b 的表达明显低于对照组。原始测序数据和处理后的数据质量均满足分析要求，miR-146b 减少后，circRNA 的表达发生了调节。其中，siRNA-miR-146b 组与对照组相比，有 18 个 circRNA 上调，77 个 circRNA 下调。基因预测表明，hsa_circ1887 是 miRNA 和 circRNA 调控网络中的最大作用点。qRT-PCR 显示，与对照组相比，siRNA-miR-146b 组 rno-circRNA-469、rno-circRNA-1138、rno-circRNA-2168 和 rno-circRNA-1907 明显降低，而 circRNA-1984 明显升高，与高通量测序技术的测量结果一致。
miR-146b 可调节 HSCs 中 circRNA 的表达，可能参与肝纤维化的形成和发展。
