Methylation Status of the miR-141-3p Promoter Regulates miR-141-3p Expression, Inflammasome Formation, and the Invasiveness of HTR-8/SVneo Cells

Cytogenet Genome Res. 2021;161(10-11):501-513. doi: 10.1159/000519740. Epub 2021 Dec 8.

Abstract

MicroRNA-141 (miR-141-3p) is upregulated in preeclampsia. This study investigated the effect of methylation of the miR-141-3p promoter on cell viability, invasion capability, and inflammasomes in vitro. The expression of miR-141-3p and methylation status of the miR-141-3p promoter were examined by RT-qPCR and pyrosequencing in villus tissues of women with spontaneous delivery (VTsd), villus tissues of women with preeclampsia (VTpe), and also in HTR-8/SVneo cells treated with a miR-141-3p inhibitor and 20 μmol/L 5-aza-2'-deoxycytidine (5-Aza), a DNA methyltransferase inhibitor. Cell viability and invasion were evaluated by CCK-8 and transwell assays. In addition, the levels of CXCL12, CXCR4, CXCR2, MMPs, NLRP3, and ASC expression were assessed by western blotting, and IL-1β and IL-18 concentrations were assayed by ELISA. miR-141-3p expression was upregulated, and the levels of miR-141-3p promoter methylation and CXCL12, CXCR4, and CXCR2 expression were decreased in VTpe relative to VTsd. In HTR-8/SVneo cells, hypomethylation caused by 5-Aza treatment increased miR-141-3p expression, while DNA methyltransferase 3 (DNMT3) transfection decreased miR-141-3p expression. miRNA-141-3p induced NLRP3, IL-1β, and IL-18 production, decreased CXCR4, MMP, and MMP2 production, and suppressed cell growth and invasion. Furthermore, we observed that NLRP3 plays an important mediatory role in the effects of miR-141-3p described above. Decreased methylation of the miR-141-3p promoter increases miR-141-3p expression, which in turn increases NLRP3 expression, resulting in higher IL-1β and IL-18 levels and lower levels of MMP2/9 and CXCR4. We conclude that modification of the miR-141-3p promoter might be a curial mediator in preeclampsia.

Keywords: Invasion; Methylation; Preeclampsia; Proliferation; microRNA-141.

MeSH terms

  • Cell Division / drug effects
  • Cell Movement / drug effects
  • Chorionic Villi / metabolism
  • Chorionic Villi / pathology
  • DNA (Cytosine-5-)-Methyltransferases / antagonists & inhibitors
  • DNA (Cytosine-5-)-Methyltransferases / metabolism
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism
  • DNA Methylation*
  • DNA Methyltransferase 3B
  • Decitabine / pharmacology
  • Female
  • Humans
  • Inflammasomes / metabolism*
  • Interleukin-18 / analysis
  • Interleukin-1beta / analysis
  • Matrix Metalloproteinases / metabolism
  • MicroRNAs / antagonists & inhibitors
  • MicroRNAs / biosynthesis
  • MicroRNAs / genetics*
  • Pre-Eclampsia / genetics
  • Pre-Eclampsia / metabolism
  • Pre-Eclampsia / pathology*
  • Pregnancy
  • Promoter Regions, Genetic / genetics*

Substances

  • Inflammasomes
  • Interleukin-18
  • Interleukin-1beta
  • MIRN141 microRNA, human
  • MicroRNAs
  • Decitabine
  • DNA
  • DNA (Cytosine-5-)-Methyltransferases
  • Matrix Metalloproteinases