Department of Medical Genetics and Prenatal Diagnosis, Lianyungang Maternal and Child Health Hospital.
Department of Pregnancy Health Management, Lianyungang Maternal and Child Health Hospital.
Biol Pharm Bull. 2021;44(4):507-514. doi: 10.1248/bpb.b20-00755.
Preeclampsia (PE) is a severe pregnancy-specific complication responsible for a majority of maternal and fetal mortality. The dysfunction of trophoblast cells is known to be associated with the etiology of PE. Moreover, elevated expression of hsa_circ_0001326 was found in patients with PE without elucidating specific mechanisms. Thus, this study aimed to investigate the role of hsa_circ_0001326 in the dysfunction of trophoblast cells in vitro. Human trophoblast SWAN71 cells were used in this study. Cell proliferation, apoptosis and cell cycle were detected by 5-ethynyl-2'-deoxyuridine (EdU) staining, cell counting kit-8 assay, Annexin V/propidium iodide (PI) staining and flow cytometry, respectively. Dual luciferase assay was performed to validate the predicted targets. Additionally, Western blot was conducted for protein detection. The results indicated overexpression (OE) of hsa_circ_0001326 remarkably decreased the viability and proliferation of SWAN71 cells. MiR-186-5p was identified as the target of hsa_circ_0001326. Meanwhile, p27 Kip1 was validated as the target of hsa_miR-186-5p. Moreover, the increased apoptosis and decreased migration induced by hsa_circ_0001326 OE were inhibited by p27 Kip1 knockdown. Hsa_circ_0001326 OE upregulated p27 Kip1 and cleaved caspase3 and downregulated CDK2 and cyclin E1 in cells, while these phenomena were reversed by p27 Kip1 knockdown. In addition, hsa_circ_0001326 OE induced G0/G1 cell cycle arrest was also attenuated in the presence of p27 Kip1 knockdown. Taken together, hsa_circ_0001326 OE contributed to the decreased viability of SWAN71 cells by targeting miR-186-5p via upregulation of p27 Kip1. Our findings were helpful to uncover the pathophysiological process of PE, as well as inspire the development of novel targeted therapy against PE.
子痫前期 (PE) 是一种严重的妊娠特异性并发症，是导致母婴死亡的主要原因。已知滋养细胞功能障碍与 PE 的病因有关。此外，在没有阐明具体机制的情况下，发现 PE 患者中 hsa_circ_0001326 的表达升高。因此，本研究旨在探讨 hsa_circ_0001326 在体外滋养细胞功能障碍中的作用。本研究使用人滋养细胞 SWAN71 细胞。通过 5-乙炔基-2'-脱氧尿苷 (EdU) 染色、细胞计数试剂盒-8 测定、膜联蛋白 V/碘化丙啶 (PI) 染色和流式细胞术分别检测细胞增殖、细胞凋亡和细胞周期。双荧光素酶报告基因实验验证预测靶点。此外，进行 Western blot 检测蛋白表达。结果表明，hsa_circ_0001326 的过表达 (OE) 显著降低了 SWAN71 细胞的活力和增殖。miR-186-5p 被鉴定为 hsa_circ_0001326 的靶标。同时，p27 Kip1 被验证为 hsa_miR-186-5p 的靶标。此外，hsa_circ_0001326 OE 诱导的细胞凋亡增加和迁移减少可被 p27 Kip1 敲低所抑制。hsa_circ_0001326 OE 上调细胞中 p27 Kip1 和裂解 caspase3，下调 CDK2 和细胞周期蛋白 E1，而这些现象可被 p27 Kip1 敲低所逆转。此外，p27 Kip1 敲低减弱了 hsa_circ_0001326 OE 诱导的 G0/G1 细胞周期阻滞。综上所述，hsa_circ_0001326 通过靶向 miR-186-5p 上调 p27 Kip1 导致 SWAN71 细胞活力降低。我们的研究结果有助于揭示 PE 的病理生理过程，并为开发针对 PE 的新型靶向治疗方法提供启示。
