Department of Pediatrics, Integrated Chinese and Western Medicine Hospital of Zhejiang Province, Hangzhou, People's Republic of China.
Department of Pediatrics, Zhejiang University Hospital, Hangzhou, People's Republic of China.
J Obstet Gynaecol Res. 2020 Jun;46(6):864-875. doi: 10.1111/jog.14249. Epub 2020 Mar 22.
High-expressed miR-330-3p in gestational diabetes mellitus (GDM) patients was reported. However, the role and mechanism of miR-330-3p in GDM are rarely reported. In this research, we aim to investigate the effects of miR-330-3p on GDM.
MiR-330-3p expression in the GDM patients' blood was determined by q-PCR. Blood glucose of blood samples was detected using blood glucose detection kits. Glucokinase (GCK) was confirmed to be a target gene of miR-330-3p by bioinformatics and luciferase analysis. Correlations between miR-330-3p with GCK and blood glucose were analyzed by Pearson correlation analysis. After INS-1 cells were treated with glucose and transfected with mimic, inhibitor or siGCK, GCK expression was detected by western blot, and q-PCR, enzyme-linked immunosorbent assays, cell counting kit-8 and Annexin-V/propidium iodide were conducted to examine the expression of insulin, cell viability and apoptosis.
MiR-330-3p was high-expressed in GDM patients' blood, while GCK was low-expressed. The miR-330-3p expression level positively correlated with blood glucoseand and it was highly expressed in glucose-treated INS-1 cells (11 and 22 mmol/L), while miR-330-3p expression negatively correlated with GCK expression. GCK expression was inhibited by miR-330-3p mimic and enhanced by the miR-330-3p inhibitor. MiR-330-3p mimic inhibited INS-1 cells' insulin expression, cell viability and induced apoptosis. Yet miR-330-3p inhibitor and siGCK exhibited opposite effects which miR-330-3p mimic and GCK played on INS-1 cells. In addition, siGCK reversed the effect of miR-330-3p inhibitor on INS-1 cells.
Our findings proved that miR-330-3p targeting GCK lead to the dysfunction of INS-1 cells in GDM, and could become a therapeutic target for GDM treatment.
有研究报道妊娠期糖尿病（GDM）患者中高表达 miR-330-3p。然而，miR-330-3p 在 GDM 中的作用和机制很少有报道。在这项研究中，我们旨在研究 miR-330-3p 对 GDM 的影响。
通过 q-PCR 检测 GDM 患者血液中的 miR-330-3p 表达。使用血糖检测试剂盒检测血样中的血糖。通过生物信息学和荧光素酶分析证实葡萄糖激酶（GCK）是 miR-330-3p 的靶基因。通过 Pearson 相关性分析分析 miR-330-3p 与 GCK 和血糖之间的相关性。用葡萄糖处理 INS-1 细胞并转染 mimics、inhibitor 或 siGCK 后，通过 Western blot、q-PCR、酶联免疫吸附测定、细胞计数试剂盒-8 和 Annexin-V/碘化丙啶检测胰岛素表达、细胞活力和细胞凋亡。
miR-330-3p 在 GDM 患者的血液中高表达，而 GCK 低表达。miR-330-3p 的表达水平与血糖呈正相关，在 11 和 22mmol/L 葡萄糖处理的 INS-1 细胞中高表达，而 miR-330-3p 的表达与 GCK 表达呈负相关。miR-330-3p mimics 抑制 GCK 表达，miR-330-3p 抑制剂增强 GCK 表达。miR-330-3p mimics 抑制 INS-1 细胞胰岛素表达、细胞活力并诱导细胞凋亡。然而，miR-330-3p 抑制剂和 siGCK 对 INS-1 细胞表现出相反的作用，miR-330-3p mimic 和 GCK 对 INS-1 细胞也表现出相反的作用。此外，siGCK 逆转了 miR-330-3p 抑制剂对 INS-1 细胞的作用。
我们的研究结果证明，miR-330-3p 靶向 GCK 导致 GDM 中 INS-1 细胞功能障碍，可能成为 GDM 治疗的一个治疗靶点。
