Department of Nephropathy, Central South University Xiangya School of Medicine Affiliated Haikou Hospital, Haikou, Hainan, China.
Department of Central Lab, Central South University Xiangya School of Medicine Affiliated Haikou Hospital, Haikou, Hainan, China.
Biomed Pharmacother. 2020 Jan;121:109622. doi: 10.1016/j.biopha.2019.109622. Epub 2019 Nov 25.
Previous study has demonstrated that long noncoding RNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) was abnormally expressed in diabetic nephropathy (DN). However, the underlying mechanism that allows CDKN2B-AS1 in the progression of DN remains to be further elucidated.
Peripheral blood cells of 24 diabetes patients with DN and 20 without DN were collected. Human glomerular mesangial cells (HGMC) were cultured in high glucose or low glucose medium. The expression levels of CDKN2B-AS1, microRNA (miR)-424-5p and high mobility group AT hook 2 (HMGA2) were detected by quantitative real-time polymerase chain reaction or western blot. The target association between miR-424-5p and CDKN2B-AS1 or HMGA2 was confirmed by dual-luciferase reporter and RNA immunoprecipitation assays. Cell proliferation, extracellular matrix (ECM) accumulation and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) and western blot, respectively.
CDKN2B-AS1 expression was up-regulated and miR-424-5p level was down-regulated in peripheral blood of DN patients and high glucose-treated HGMC cells. CDKN2B-AS1 was validated as a sponge of miR-424-5p. Silence of CDKN2B-AS1 repressed proliferation and ECM accumulation by increasing miR-424-5p. HMGA2 was a target of miR-424-5p and miR-424-5p overexpression inhibited proliferation, ECM accumulation and PI3K/AKT pathway by targeting HMGA2. Moreover, knockdown of CDKN2B-AS1 inhibited HMGA2 expression and PI3K/AKT pathway by increasing miR-424-5p.
Knockdown of CDKN2B-AS1 suppressed proliferation, ECM accumulation and PI3K/AKT signaling by increasing miR-424-5p and decreasing HMGA2 in high glucose-treated HMGC cells.
先前的研究表明，长链非编码 RNA 细胞周期蛋白依赖性激酶抑制剂 2B 反义 RNA 1（CDKN2B-AS1）在糖尿病肾病（DN）中异常表达。然而，CDKN2B-AS1 在 DN 进展中的潜在机制仍有待进一步阐明。
收集 24 例糖尿病合并 DN 患者和 20 例无 DN 患者的外周血细胞，培养高糖或低糖条件下人肾小球系膜细胞（HGMC）。采用实时定量聚合酶链反应或 Western blot 检测 CDKN2B-AS1、微小 RNA（miR）-424-5p 和高迁移率族 AT 钩 2（HMGA2）的表达水平。通过双荧光素酶报告和 RNA 免疫沉淀实验证实 miR-424-5p 与 CDKN2B-AS1 或 HMGA2 的靶基因结合。通过 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴盐（MTT）和 Western blot 分别检测细胞增殖、细胞外基质（ECM）积累和磷脂酰肌醇 3-激酶（PI3K）/蛋白激酶 B（AKT）信号通路。
DN 患者外周血和高糖处理的 HGMC 细胞中 CDKN2B-AS1 表达上调，miR-424-5p 水平下调。CDKN2B-AS1 被证实是 miR-424-5p 的海绵。沉默 CDKN2B-AS1 通过增加 miR-424-5p 抑制增殖和 ECM 积累。HMGA2 是 miR-424-5p 的靶基因，miR-424-5p 过表达通过靶向 HMGA2 抑制增殖、ECM 积累和 PI3K/AKT 通路。此外，沉默 CDKN2B-AS1 通过增加 miR-424-5p 抑制 HMGA2 表达和 PI3K/AKT 通路。
沉默 CDKN2B-AS1 通过增加 miR-424-5p 抑制高糖处理的 HGMC 细胞中 HMGA2 的表达和 PI3K/AKT 信号通路，抑制细胞增殖、ECM 积累和 PI3K/AKT 信号通路。
