Abstract：
Fetal growth restriction (FGR) severely affects the health outcome of newborns and represents a major cause of perinatal morbidity. The precise involvement of circCULT1 in the progression of FGR remains unclear. We performed next-generation sequencing and RT-qPCR to identify differentially expressed circRNAs in placental tissues affected by FGR by comparing them with unaffected counterparts. Edu, flow cytometry, and transwell assay were conducted to detect HTR8/SVneo cell\'s function in regard to cell proliferation, migration, and invasion. The interaction between circCUL1 and hsa-miR-30e-3p was assessed through dual-luciferase reporter assays, validation of the interaction between circCUL1 and ANXA1 was performed using RNA pulldown and immunoprecipitation assays. Western blot analysis was performed to evaluate protein levels of autophagy markers and components of the PI3K/AKT signaling pathway. A knockout (KO) mouse model was established for homologous mmu-circ-0001469 to assess fetal mouse growth and development indicators. Our findings revealed an upregulation of circCUL1 expression in placental tissues from patients with FGR. We found that suppression of circCUL1 increased the trophoblast cell proliferation, migration, and invasion, circCUL1 could interact with hsa-miR-30e-3p. Further, circCUL1 stimulated autophagy, modulating trophoblast cell autophagy via the ANXA1/PI3K/AKT pathway, and a notable disparity was observed, with KO mice displaying accelerated embryo development and exhibiting heavier placentas in comparison to wild-type C57BL/6 mice. By modulating the ANXA1/PI3K/AKT signaling pathway through the interaction with hsa-miR-30e-3p, circCUL1 promotes autophagy while concurrently suppressing trophoblast cell proliferation, migration, and invasion. These findings offer novel insights into potential diagnostic markers and therapeutic targets for FGR research.
摘要：
胎儿生长受限（FGR）严重影响新生儿的健康结局，是围产期发病的主要原因。circULT1在FGR进展中的确切参与尚不清楚。我们进行了下一代测序和RT-qPCR，以通过将其与未受影响的对应物进行比较来鉴定受FGR影响的胎盘组织中差异表达的circRNAs。Edu,流式细胞术,和transwell试验进行检测HTR8/SVneo细胞在细胞增殖方面的功能，迁移,和入侵。circUL1和hsa-miR-30e-3p之间的相互作用通过双荧光素酶报告基因测定进行评估，circUL1和ANXA1之间的相互作用的验证使用RNA下拉和免疫沉淀测定进行.进行蛋白质印迹分析以评估自噬标志物和PI3K/AKT信号通路组分的蛋白水平。建立同源mmu-circ-0001469的敲除(KO)小鼠模型以评估胎儿小鼠的生长发育指标。我们的发现揭示了FGR患者胎盘组织中circUL1表达的上调。我们发现circcUL1的抑制增加了滋养层细胞的增殖，迁移,和入侵,circUL1可以与hsa-miR-30e-3p相互作用。Further,circUL1刺激自噬，通过ANXA1/PI3K/AKT通路调节滋养层细胞自噬,观察到明显的差异，与野生型C57BL/6小鼠相比，KO小鼠表现出加速的胚胎发育并表现出较重的胎盘。通过与hsa-miR-30e-3p相互作用调节ANXA1/PI3K/AKT信号通路,circUL1促进自噬，同时抑制滋养细胞增殖，迁移,和入侵。这些发现为FGR研究提供了对潜在诊断标志物和治疗靶标的新见解。
