Introduction: Many studies have shown that circular RNAs (circRNAs) are related to the occurrence of preeclampsia (PE). However, the role of circLRRK1 in the progression of PE is unclear.
Methods: The identification and localization of circLRRK1 were verified by Actinomycin D (ActD) assay, Ribonuclease R (RNase R) digestion assay and subcellular localization assay. Moreover, the proliferation of trophoblast cells was detected by 3-(45)-dimethylthiahiazo (-z-y1)-35-di-phenytetrazoliumromide (MTT) assay and colony formation assay. Furthermore, the migration and invasion of trophoblast cells were determined by wound healing assay and transwell assay. Meanwhile, Western blot (WB) analysis was used to examine the protein levels of migration markers and PI3K/AKT signaling pathway markers. In addition, the interaction between circLRRK1 and miR-223-3p was confirmed by dual-luciferase reporter assay and biotin-labeled RNA pull-down assay.
Results: Our results showed that circLRRK1 was significantly highly expressed in PE patients. Silenced circLRRK1 could markedly enhance the proliferation, migration and invasion of trophoblast cells. Additionally, we found that circLRRK1 could target miR-223-3p. MiR-223-3p overexpression also promoted the proliferation, migration and invasion of trophoblast cells. The rescue experiments revealed that miR-223-3p inhibitor could reverse the promoting effect of circLRRK1 silencing on the proliferation, migration and invasion of trophoblast cells. Furthermore, circLRRK1 silencing could activate the PI3K/AKT signaling pathway by targeting miR-223-3p.
Discussion: CircLRRK1 could suppress the proliferation, migration and invasion of trophoblast cells by regulating the PI3K/AKT signaling pathway via targeting miR-223-3p, suggesting that circLRRK1 might be a potential biomarker for the treatment of PE.
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