Let-7a inhibits Bcl-xl and YAP1 expression to induce apoptosis of trophoblast cells in early-onset severe preeclampsia

Abstract

Dysregulation of the MicroRNA (miR) Let-7 family has been implicated in preeclampsia (PE). Abnormal trophoblast cell proliferation and apoptosis associate with the pathogenesis of PE. The present study was designed to test the hypothesis whether let-7a could regulate the biological functions of trophoblasts and explore the mechanism how it works in the development of early-onset severe PE. The putative target genes Bcl-xl and YAP1 of let-7a were verified by luciferase assay. The roles of let-7a, Bcl-xl and YAP1 in regulating JEG-3 cell functions were examined by altering their expression with mimic, overexpression plasmids or siRNAs. The methylation status of let-7a-3 in PE was assessed by methylation-specific and bisulfite sequencing PCR assays. JEG-3 cells were treated with DNA methyltransferase inhibitor to analyze whether let-7a-3 demethylation functioned in PE. Tumor growth and cell apoptosis were measured from nude mice inoculated with JEG-3 cells overexpressing let-7a. The results revealed let-7a was highly expressed in early-onset severe PE and let-7a-3 presented a low methylation level. Functionally, let-7a upregulation could inhibit the viability and cell cycle progression but induce the apoptosis of JEG-3 cells. Bcl-xl and YAP1, target genes of let-7a, could rescue cell apoptosis induced by let-7a. The demethylation of let-7a-3 was also observed to elevate the expression of let-7a and enhance JEG-3 cell apoptosis. Let-7a inhibited tumorigenic ability of JEG-3 cells and enhanced cell apoptosis in vivo. Altogether, let-7a could enhance cell apoptosis in trophoblasts through downregulation of Bcl-xl and YAP1, which suggests that let-7a might be a key regulator in the progression of PE.

Keywords: Bcl-xl; C-caspase-3; C-caspase-9; Hypomethylation; Let-7a-3; MicroRNA; Preeclampsia; Trophoblast cells; YAP1.