Department of Clinical Pharmacology, Xiangya Hospital, Central South University, 410008, Changsha, China.
Institute of Clinical Pharmacology, Central South University, Hunan Key Laboratory of Pharmacogenetics, Changsha, 410008, Hunan, China.
Endocrine. 2019 Mar;63(3):489-496. doi: 10.1007/s12020-018-1754-6. Epub 2018 Sep 21.
Diabetes mellitus is a major chronic disease and causes over one million deaths. KCNJ11 genetic polymorphisms influence the response of first-line oral antidiabetic agent sulfonylureas. Hsa-miR-4532 correlates with diabetic nephropathy and has a high abundance in urine. MIR4532 rs60452575 G>A variant changes the mature sequence of hsa-miR-4532. We studied whether the genetic polymorphisms of MIR4532 rs60452575 would influence KCNJ11 expression and sulfonylurea-stimulated insulin secretion or not.
To estimate the influence that rs60452575 G>A variant has on the interaction of hsa-miR-4532 and KCNJ11, we constructed a pmirGLO vector containing 3' UTR of KCNJ11 and co-transfected it with wild-type and mutant hsa-miR-4532 mimics into HEK293 cells; and we overexpressed wild-type and mutant hsa-miR-4532 mimics into HEK293 cells and MIN6 cells to access its effects on KCNJ11 expression and response of sulfonylureas.
MIR4532 rs60452575 G>A variant appeared to disrupt the repression of KCNJ11 expression in both cell lines, and reduce the sulfonylurea-stimulated insulin secretion by breaking the binding of the hsa-miR-4532 to 3' UTR of KCNJ11 in MIN6 cells.
Our study indicates that MIR4532 rs60452575 variant influences KCNJ11 expression and sulfonylurea response. It might be a potential predictive factor of sulfonylureas therapy.
糖尿病是一种主要的慢性疾病，导致超过 100 万人死亡。KCNJ11 基因多态性影响一线口服降糖药物磺酰脲类药物的反应。hsa-miR-4532 与糖尿病肾病相关，在尿液中丰度较高。MIR4532 rs60452575 G>A 变体改变了 hsa-miR-4532 的成熟序列。我们研究了 MIR4532 rs60452575 的遗传多态性是否会影响 KCNJ11 的表达和磺酰脲类药物刺激的胰岛素分泌。
为了评估 rs60452575 G>A 变体对 hsa-miR-4532 与 KCNJ11 相互作用的影响，我们构建了一个包含 KCNJ11 3'UTR 的 pmirGLO 载体，并将其与野生型和突变型 hsa-miR-4532 模拟物共转染到 HEK293 细胞中；我们过表达野生型和突变型 hsa-miR-4532 模拟物到 HEK293 细胞和 MIN6 细胞中，以评估其对 KCNJ11 表达和磺酰脲类药物反应的影响。
MIR4532 rs60452575 G>A 变体似乎在两种细胞系中破坏了 KCNJ11 表达的抑制作用，并通过破坏 hsa-miR-4532 与 MIN6 细胞中 KCNJ11 3'UTR 的结合，减少了磺酰脲类药物刺激的胰岛素分泌。
我们的研究表明，MIR4532 rs60452575 变体影响 KCNJ11 表达和磺酰脲类药物反应。它可能是磺酰脲类药物治疗的潜在预测因子。
