Department of Emergency, The First Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang, 421001, Hunan, China.
Department of Dermatology, The First Affiliated Hospital, Hengyang Medical School, University of South China, No. 69, Chuanshan Road, Hengyang, 421001, Hunan, China.
Arch Dermatol Res. 2023 Sep;315(7):2079-2090. doi: 10.1007/s00403-023-02597-3. Epub 2023 Mar 21.
Circular RNA (circRNA) has been confirmed to participate in psoriasis process, but the role of circ_0024028 in psoriasis development is still unclear. Interleukin 22 (IL-22)-induced keratinocytes (HaCaT) were used to construct psoriasis cell models in vitro. The expression of circ_0024028, microRNA (miR)-486-3p and AKT serine/threonine kinase 3 (AKT3) was analyzed by quantitative real-time PCR. Cell function was assessed by cell counting kit 8 assay, EdU assay, transwell assay, and wound healing assay. Protein expression was examined using western blot analysis. RNA interaction was confirmed by dual-luciferase reporter assay and RIP assay. Exosomes were isolated from cell culture medium using ultracentrifugation and examined by transmission electron microscopy and nanoparticle tracking analysis. Circ_0024028 was highly expressed in psoriasis lesions and IL-22-induced HaCaT cells, and its silencing could inhibit IL-22-induced HaCaT cell proliferation and migration. MiR-486-3p could be sponged by circ_0024028, and its inhibitor restored the functions of circ_0024028 knockdown on IL-22-induced HaCaT cell proliferation and migration. AKT3 was targeted by miR-486-3p, and its overexpression reversed the inhibitory effect of miR-486-3p on IL-22-induced HaCaT cell proliferation and migration. AKT3 expression was positively regulated by circ_0024028, and circ_0024028/miR-486-3p/AKT3 axis could regulate the activity of AKT/mTOR pathway. Additionally, exosomes mediated the transfer of circ_0024028 in cells. Circ_0024028 might be a potential target for psoriasis treatment, which knockdown repressed IL-22-induced keratinocytes proliferation and migration through miR-486-3p/AKT3 pathway.
环状 RNA(circRNA)已被证实参与银屑病的发生过程,但 circ_0024028 在银屑病发病中的作用尚不清楚。本研究采用白细胞介素 22(IL-22)诱导角质形成细胞(HaCaT)构建体外银屑病细胞模型,采用实时定量 PCR 分析 circ_0024028、微小 RNA(miR)-486-3p 和丝氨酸/苏氨酸蛋白激酶 3(AKT3)的表达。通过细胞计数试剂盒 8 检测、EdU 检测、Transwell 检测和划痕愈合试验评估细胞功能。采用 Western blot 检测蛋白表达。通过双荧光素酶报告基因检测和 RNA 免疫沉淀(RIP)实验验证 RNA 相互作用。采用超速离心法从细胞培养液中分离外泌体,通过透射电子显微镜和纳米颗粒跟踪分析进行检测。circ_0024028 在银屑病皮损和 IL-22 诱导的 HaCaT 细胞中高表达,其沉默可抑制 IL-22 诱导的 HaCaT 细胞增殖和迁移。circ_0024028 可作为 miR-486-3p 的海绵,miR-486-3p 的抑制剂可恢复 circ_0024028 敲低对 IL-22 诱导的 HaCaT 细胞增殖和迁移的作用。AKT3 是 miR-486-3p 的靶基因,其过表达可逆转 miR-486-3p 对 IL-22 诱导的 HaCaT 细胞增殖和迁移的抑制作用。circ_0024028 对 AKT3 的表达具有正向调控作用,circ_0024028/miR-486-3p/AKT3 轴可调节 AKT/mTOR 通路的活性。此外,外泌体介导了 circ_0024028 在细胞间的转移。circ_0024028 可能是治疗银屑病的潜在靶点,其敲低通过 miR-486-3p/AKT3 通路抑制 IL-22 诱导的角质形成细胞增殖和迁移。