Department of Obstetrics, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Int J Mol Med. 2021 Jun;47(6). doi: 10.3892/ijmm.2021.4944. Epub 2021 Apr 28.
Circular (circ)RNA has been demonstrated to serve crucial roles in cell proliferation, differentiation and autophagy. However, to date, the function and mechanism of action of circRNA in preeclampsia have not been reported. The present study aimed to analyze the roles of circRNA‑0004904 in preeclampsia and to clarify its underlying pathogenic mechanism. The expression levels of circ‑0004904, microRNA (miR)‑570 and autophagy‑related 12 (ATG12) were detected by reverse transcription‑quantitative (RT‑q)PCR. In addition, the protein levels of ATG12, vascular endothelial growth factor (VEGF) and fused in sarcoma (FUS) were determined by western blot assay. The distribution of mRFP‑GFP‑LC3 in HTR8 and JEG3 cells was analyzed by confocal microscopy. Fluorescence hybridization assay was utilized to identify the colocalization of circ‑0004904 and miR‑570. Cell proliferation was determined by 5‑ethynyl‑2'‑deoxyuridine assay, and invasion was evaluated by Matrigel invasion assay. The results of the present study demonstrated that the expression levels of circ‑0004904 were elevated in the placental tissues and plasma samples of patients with preeclampsia compared with those in the control group samples. Ectopic expression of circ‑0004904 promoted autophagy, but inhibited migration and proliferation of HTR8 cells compared with those in the negative control group. Silencing of circ‑0004904 inhibited autophagy, and induced migration and proliferation in JEG3 cells compared with those in the negative control group. In addition, circ‑0004904 regulated the levels of ATG12 via interaction with miR‑570. Furthermore, circ‑0004904 regulated the FUS/VEGF axis in HTR8 and JEG3 cells. In conclusion, circ‑0004904 was abnormally expressed in the plasma and placental tissues of patients with preeclampsia. In addition, circ‑0004904 was involved in the regulation of proliferation, invasion and autophagy in HTR8 and JEG3 cells. Thus, circ‑0004904 may be used as a potential diagnostic biomarker and therapeutic target for preeclampsia.
环状 RNA（circRNA）已被证明在细胞增殖、分化和自噬中发挥关键作用。然而，迄今为止，circRNA 在子痫前期中的作用及其作用机制尚不清楚。本研究旨在分析 circRNA-0004904 在子痫前期中的作用，并阐明其潜在的发病机制。通过逆转录-定量（RT-q）PCR 检测 circ-0004904、微小 RNA（miR）-570 和自噬相关 12（ATG12）的表达水平。此外，通过 Western blot 检测 ATG12、血管内皮生长因子（VEGF）和融合肉瘤（FUS）的蛋白水平。通过共聚焦显微镜分析 HTR8 和 JEG3 细胞中 mRFP-GFP-LC3 的分布。荧光杂交试验用于鉴定 circ-0004904 和 miR-570 的共定位。通过 5-乙炔基-2'-脱氧尿苷测定法测定细胞增殖，通过 Matrigel 侵袭测定法评估侵袭。本研究结果表明，与对照组样本相比，子痫前期患者胎盘组织和血浆样本中 circ-0004904 的表达水平升高。与阴性对照组相比，外源性表达 circ-0004904 促进 HTR8 细胞自噬，但抑制其迁移和增殖。与阴性对照组相比，沉默 circ-0004904 抑制 JEG3 细胞自噬，并诱导其迁移和增殖。此外，circ-0004904 通过与 miR-570 相互作用调节 ATG12 的水平。此外，circ-0004904 调节 HTR8 和 JEG3 细胞中的 FUS/VEGF 轴。综上所述，circ-0004904 在子痫前期患者的血浆和胎盘组织中异常表达。此外，circ-0004904 参与调节 HTR8 和 JEG3 细胞的增殖、侵袭和自噬。因此，circ-0004904 可用作子痫前期的潜在诊断生物标志物和治疗靶点。
