More and more evidence has identified that long non-coding RNAs (lncRNAs) are involved in various biological process of numerous diseases. It has been reported that long intergenic non-protein coding RNA 473 (LINC00473) was associated with pre-eclampsia (PE) development. However, role and molecular mechanism of LINC00473 in PE remains elusive. Therefore, we designed this research to figure out the specific biological function of LINC00473 in trophoblasts. Firstly, we testified expressions of LINC00473 in trophoblasts of PE with RT-qPCR analysis. Then, to probe biological function of LINC00473 in trophoblasts of PE, CCK-8 assay, trans-well assays and western blot analysis were conducted in Wish and JAR cells. As for verifying interaction of microRNA-15a-5p (miR-15a-5p) and LINC00473 or lipopolysaccharide induced TNF factor (LITAF), RNA pull-down and luciferase reporter assays were carried out. Finally, rescue experiments were conducted to probe regulatory pattern of the LINC00473/miR-15a-5p/LITAF axis in trophoblasts of PE. As a result, LINC00473 presented a significant upregulation in trophoblasts of PE. Moreover, LINC00473 knockdown induced trophoblast viability, migration, invasion, and epithelial-to-mesenchymal transition (EMT) in trophoblasts. Additionally, miR-15a-5p interacted with LINC00473 and miR-15a-5p was negatively regulated by LINC00473 in trophoblasts. Simultaneously, miR-15a-5p negatively modulated LITAF in trophoblasts. Moreover, LITAF overexpression or miR-15a-5p downregulation reversed the promotive impact of silenced LINC00473 on trophoblast viability, migration, invasion and EMT. In conclusion, LINC00473 regulated migration and invasion in trophoblasts via the miR-15a-5p/LITAF axis. Our study may provide a novel insight for clinical treatment of PE.

中文翻译：

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LINC00473 下调通过 miR-15a-5p/LITAF 轴促进子痫前期滋养层细胞迁移和侵袭

越来越多的证据表明，长链非编码RNA（lncRNA）参与了多种疾病的各种生物学过程。据报道，长基因间非蛋白编码 RNA 473 (LINC00473) 与先兆子痫 (PE) 发展有关。然而，LINC00473 在 PE 中的作用和分子机制仍然难以捉摸。因此，我们设计了这项研究来弄清楚 LINC00473 在滋养层中的特定生物学功能。首先，我们通过 RT-qPCR 分析验证了 LINC00473 在 PE 滋养层中的表达。然后，为了探索 LINC00473 在 PE 滋养层中的生物学功能，在 Wish 和 JAR 细胞中进行了 CCK-8 测定、跨孔测定和蛋白质印迹分析。至于验证 microRNA-15a-5p (miR-15a-5p) 和 LINC00473 或脂多糖诱导的 TNF 因子 (LITAF) 的相互作用，进行了 RNA 下拉和荧光素酶报告基因分析。最后，进行救援实验以探讨 LINC00473/miR-15a-5p/LITAF 轴在 PE 滋养层中的调控模式。结果，LINC00473 在 PE 的滋养层中表现出显着的上调。此外，LINC00473 敲低可诱导滋养细胞中的滋养细胞活力、迁移、侵袭和上皮间质转化 (EMT)。此外，miR-15a-5p 与 LINC00473 相互作用，并且 miR-15a-5p 在滋养细胞中受到 LINC00473 的负调控。同时，miR-15a-5p 在滋养细胞中负调节 LITAF。此外，LITAF 过表达或 miR-15a-5p 下调逆转了沉默的 LINC00473 对滋养层活力、迁移、侵袭和 EMT 的促进作用。综上所述，LINC00473 通过 miR-15a-5p/LITAF 轴调节滋养细胞的迁移和侵袭。我们的研究可能为 PE 的临床治疗提供新的见解。