The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Ren Fail. 2023 Dec;45(1):2204953. doi: 10.1080/0886022X.2023.2204953.
Recent evidence suggested that long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) play critical roles in the pathogenesis of vascular calcification (VC). In this study, we tried to explore the expression and role of a lncRNA, i.e., metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), and a miRNA, i.e., miR-30c, in VC.
VC model was induced in human vascular smooth muscle cells (VSMCs) after 10 days culture in calcifying medium containing 2 mM NaHPO. Alizarin red S staining, calcium assay and western blot analysis of runt-related transcription factor 2 (Runx2) and alpha smooth muscle actin (α-SMA) were performed to evaluate VC. Knockdown of MALAT1 and up-regulation of MALAT1, miR-30c and Runx2 was performed to determine the impact of these molecules on VSMCs calcification. Dual-luciferase report assay was performed to confirm the relationship between MALAT1 and miR-30c or miR-30c and Runx2. In addition, quantitative reverse transcription PCR and western blot were used to determine gene and protein expression.
MALAT1 was increased, while miR-30c was decreased in calcified VSMCs. Knockdown of MALAT1 suppressed VSMCs calcification; on the contrary, up-regulation of MALAT1 promoted VSMCs calcification. The effect of MALAT1 over-expression on VSMCs calcification was reversed by upregulation of miR-30c, which was reversed again by upregulation of Runx2. Dual-luciferase report assay confirmed that there is a direct interaction between MALAT1 and miR-30c, and Runx2 is a direct target of miR-30c.
MALAT1 over-expression promoted VSMCs calcification, which was at least partially through regulating the miR-30c/Runx2 axis.
最近的证据表明，长链非编码 RNA（lncRNA）和 microRNA（miRNA）在血管钙化（VC）的发病机制中发挥着关键作用。在这项研究中，我们试图探讨 lncRNA，即转移相关肺腺癌转录物 1（MALAT1）和 miRNA，即 miR-30c 在 VC 中的表达和作用。
在含有 2mM NaHPO 的钙化培养基中培养 10 天后，诱导人血管平滑肌细胞（VSMCs）形成 VC 模型。采用茜素红 S 染色、钙测定和 runt 相关转录因子 2（Runx2）和α平滑肌肌动蛋白（α-SMA）的 Western blot 分析来评估 VC。通过 MALAT1 敲低和 MALAT1、miR-30c 和 Runx2 的上调来确定这些分子对 VSMCs 钙化的影响。双荧光素酶报告实验证实 MALAT1 与 miR-30c 或 miR-30c 与 Runx2 之间的关系。此外，采用定量逆转录 PCR 和 Western blot 法检测基因和蛋白表达。
在钙化的 VSMCs 中，MALAT1 增加，而 miR-30c 减少。MALAT1 敲低抑制 VSMCs 钙化；相反，上调 MALAT1 促进 VSMCs 钙化。上调 miR-30c 逆转了 MALAT1 过表达对 VSMCs 钙化的影响，而上调 Runx2 再次逆转了这一影响。双荧光素酶报告实验证实 MALAT1 与 miR-30c 之间存在直接相互作用，而 Runx2 是 miR-30c 的直接靶标。
MALAT1 过表达促进了 VSMCs 的钙化，这至少部分是通过调节 miR-30c/Runx2 轴实现的。
