Cross-talk between miR-29 and Transforming Growth Factor-Betas in Trabecular Meshwork Cells
Cross-talk between miR-29 and Transforming Growth Factor-Betas in Trabecular Meshwork Cells
复制标题
DOI：
10.1167/iovs.10-6448
复制
发表时间：
2011-05-01
影响因子：
4.4
通讯作者：
Gonzalez, Pedro
中科院分区：
文献类型：
--
作者：
Luna, Coralia;Li, Guorong;Gonzalez, Pedro
目的。研究细胞外基质 (ECM) 负调节因子 microRNA-29 (miR-29) 与转化生长因子 (TGF)beta-1 和 TGF beta-2 之间的相互作用。方法。用 TGF beta 1 和 TGF beta 2 (1 ng/mL) 处理后，通过定量 PCR (Q-PCR) 分析 miR-29 家族表达的变化。在用 miR-29b 或乱序对照转染的人小梁网 (HTM) 细胞中，分别通过 Q-PCR 和 ELISA 评估 TGF beta 1 和 TGF beta 2 的基因表达和蛋白质水平。使用腺病毒和报告基因 SEAP 分析 TGF beta 1 启动子活性。在转染miR-29b或乱序对照并用TGFβ2处理的细胞中评估miR-29b和TGFβ2对ECM基因表达的影响，并通过Q-PCR分析ECM基因的表达。结果。 TGF beta 2 但不是 TGF beta 1 下调了 miR-29 家族的三个成员。 miR-29b 的过表达可以拮抗 TGF beta 2 对几种 ECM 成分表达的影响。 MiR-29b 在启动子、转录本和蛋白水平上降低 TGF beta 1 的表达，但对活性 TGF beta 2 的表达只有很小的影响。与表达质粒的骨形态发生蛋白 1 共转染后，miR-29b 对 TGF beta 1 的抑制部分恢复。结论。结果显示 TGF beta 和 miR-29 之间存在一定程度的串扰。具体而言，TGF beta 2 对 miR-29 的下调有助于该细胞因子在 TM 细胞中诱导多种 ECM 成分。这一观察结果，连同 miR-29b 对 TGF beta 1 表达的抑制作用，表明 miR-29 家族可能在调节 TGF beta 流出途径中发挥重要作用。 （投资眼科可见科学。2011；52：3567-3572）DOI：10.1167/iovs.10-6448
PURPOSE. To investigate the interactions between microRNA-29 (miR-29), a negative regulator of extracellular matrix (ECM), and transforming growth factors (TGF)beta-1 and TGF beta-2.METHODS. Changes in expression of the miR-29 family were analyzed by quantitative-PCR (Q-PCR) after treatment with TGF beta 1 and TGF beta 2 (1 ng/mL). TGF beta 1 and TGF beta 2 were evaluated at gene expression and protein levels by Q-PCR and ELISA, respectively, in human trabecular meshwork (HTM) cells transfected with miR-29b or scramble control. TGF beta 1 promoter activity was analyzed using an adenovirus with the reporter SEAP. The effects of miR-29b and TGF beta 2 on ECM gene expression were evaluated in cells transfected with miR-29b or scramble control and treated with TGF beta 2, and the expression of ECM genes was analyzed by Q-PCR.RESULTS. TGF beta 2 but not TGF beta 1, downregulated the three members of the miR-29 family. Overexpression of miR-29b antagonized the effects of TGF beta 2 on the expression of several ECM components. MiR-29b decreased the expression of TGF beta 1 at the promoter, transcript, and protein levels but had only a minor effect on the expression of active TGF beta 2. The inhibition of TGF beta 1 by miR-29b was partially recovered after co-transfection with a plasmid-expressing bone morphogenetic protein 1.CONCLUSIONS. Results showed some level of crosstalk between TGF beta s and miR-29. Specifically, the downregulation of miR-29 by TGF beta 2 contributed to the induction of several ECM components by this cytokine in TM cells. This observation, together with the inhibitory effects of miR-29b on the expression of TGF beta 1, suggests that the miR-29 family could play an important role in modulating TGF beta s on the outflow pathway. (Invest Ophthalmol Vis Sci. 2011;52:3567-3572) DOI: 10.1167/iovs.10-6448