Department of Clinical Nutrition, Yuhuangding Hospital Affiliated to Qingdao University, No. 20 East Yuhuangding Road, Yantai, 264000, Shandong, China.
Department of Obstetrics and Gynecology, Yuhuangding Hospital Affiliated to Qingdao University, No. 20 East Yuhuangding Road, Yantai, 264000, Shandong, China.
Reprod Biol Endocrinol. 2020 Jun 6;18(1):61. doi: 10.1186/s12958-020-00618-8.
Gestational diabetes mellitus (GDM) is the most common medical complication associated with pregnancy, which may impose risks on both mother and fetus. Micro RNAs (miRNAs) and long noncoding RNAs (lncRNAs) are implied as vital regulators in GDM. A recent paper revealed dysregulation of miR-33a-5p in placental tissues of GDM patients. However, the biological function of miR-33a-5p in GDM remains elusive. This study focused on exploring the function and underlying mechanisms of miR-33a-5p in GDM.
12 GDM pregnancies and 12 healthy pregnancies were enrolled in the study. INS-1 cell line was applied in in vitro experiments. The expression levels of miR-33a-5p, lnc-DANCR (Differentiation Antagonizing Non-Protein Coding RNA), and ABCA1 (ATP-binding cassette transporter 1) mRNA were determined by RT-qPCR assay. Glucose and insulin levels were measured by ELISA assay. Luciferase reporter assay and western blot assay were applied to validate the target of miR-33a-5p.
miR-33a-5p was upregulated in the blood samples from GDM, and was positively correlated with blood glucose (p < 0.0001). Overexpression or inhibition of miR-33a-5p significantly inhibited or promoted cell growth and insulin production of INS-1 cells (p < 0.01). Furthermore, ABCA1 is a direct target of miR-33a-5p, and lnc-DANCR functions as a sponge for miR-33a-5p to antagonize the function of miR-33a-5p in INS-1 cells.
Our study demonstrated that lnc-DANCR-miR-33a-5p-ABCA1 signaling cascade plays a crucial role in the regulation of the cellular function of INS-1 cells.
妊娠期糖尿病（GDM）是与妊娠相关的最常见的医学并发症，可能对母婴双方都有风险。微小 RNA（miRNA）和长非编码 RNA（lncRNA）被认为是 GDM 的重要调节因子。最近的一篇论文揭示了 GDM 患者胎盘组织中 miR-33a-5p 的失调。然而，miR-33a-5p 在 GDM 中的生物学功能仍然难以捉摸。本研究旨在探讨 miR-33a-5p 在 GDM 中的功能及其潜在机制。
本研究纳入了 12 例 GDM 妊娠和 12 例健康妊娠。应用 INS-1 细胞系进行体外实验。采用 RT-qPCR 法检测 miR-33a-5p、lnc-DANCR（分化拮抗非编码 RNA）和 ABCA1（ATP 结合盒转运体 1）mRNA 的表达水平。采用 ELISA 法检测葡萄糖和胰岛素水平。应用荧光素酶报告基因检测和 Western blot 检测验证 miR-33a-5p 的靶基因。
miR-33a-5p 在 GDM 患者的血液样本中上调，并与血糖呈正相关（p<0.0001）。miR-33a-5p 的过表达或抑制显著抑制或促进 INS-1 细胞的细胞生长和胰岛素产生（p<0.01）。此外，ABCA1 是 miR-33a-5p 的直接靶基因，lnc-DANCR 作为 miR-33a-5p 的海绵，拮抗 INS-1 细胞中 miR-33a-5p 的功能。
本研究表明，lnc-DANCR-miR-33a-5p-ABCA1 信号级联在调节 INS-1 细胞的细胞功能中起关键作用。
