ABCA1 是 miR-144-3p 在软骨细胞中的直接靶基因

Ho Thi Bich Phuong, Nguyen Dang Quan, Phan Thi Kim Tram, Tang Ha Nam Anh, Le Thi Truc Linh
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摘要

据报道,MicroRNA144-3p 与骨关节炎(OA)有关,因为它在这种疾病中上调。研究还发现 ABCA1 也与 OA 有关。生物信息学算法显示 ABCA1 是 miR-144-3p 的潜在靶点。本研究旨在通过实验证明 ABCA1 是 miR-144-3p 的直接靶标。在对软骨细胞进行 miR-144-3p 的功能增益和功能缺失实验后,用实时 RT-PCR 测定了 ABCA1 的表达。对含有 miR-144-3p 多个结合位点的 3'UTR 进行亚克隆。还创建了 miR-144-3p 结合位点突变的载体。进行荧光素酶检测以检验 miR-144-3p 与 ABCA1 结合的能力。实时 RT-PCR 显示,miR-144-3p 的过表达抑制了 ABCA1 的表达。荧光素酶试验表明,miR-144-3p 通过其在 3'UTR 上的结合位点与 ABCA1 直接相互作用。这些数据表明,ABCA1 是 miR-144-3p 的直接靶标。
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ABCA1 is direct target gene of miR-144-3p in chondrocyte
MicroRNA144-3p was reported to associate with osteoarthritis (OA) since it was upregulated in this disease. ABCA1 was also found to involve in OA. Bioinformatics algorithms showed ABCA1 was potential targets of miR-144-3p. This study aims to prove ABCA1 is direct targets of miR-144-3p experimentally. Expression of ABCA1 was determined by Realtime RT-PCR after performing the gain- and loss- function of miR-144-3p in chondrocyte. The 3’UTR containing several binding sites of miR-144-3p was subcloning. The vector with binding sites of miR-144-3p mutated was aslo created. Luciferase assay was performed to check the ability of miR-144-3p binding to ABCA1. Realtime RT-PCR showed that the overexpression of miR-144-3p inhibited ABCA1 expression. The Luciferase assay showed that miR-144-3p directly interacted with ABCA1 through its binding sites on the 3’UTR. These data suggested that ABCA1 is the direct target of miR-144-3p.
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