Recurrent spontaneous abortion (RSA) can be attributed to multiple factors, and abnormal invasion and apoptosis of trophoblast cells have attracted extensive attention. Our study aimed to investigate the expression of PTEN and miRNAs with potential regulatory relationships in the placental villi of RSA patients. Nineteen RSA patients and sixteen healthy women at reproductive age undergoing induced abortion (IA) were enrolled in the present study. The expression of PTEN and miRNAs were investigated using real-time polymerase chain reaction (PCR) and Western blotting, further verification between PTEN and potential miRNAs used cell culture and transfection, and luciferase activity assays were used to determine whether PTEN is directly regulated by potential miRNA. The results indicated that both PTEN mRNA and protein expression levels were upregulated in RSA patients, but a significant difference was only observed in protein expression level (p < 0.001). Through real-time PCR pre-scanning, the results of nine potential miRNAs revealed three significantly upregulated miRNAs (miR-494, miR-146a, and miR-21) and one significantly downregulated miRNA (miR-19b). The results of further verification regarding miR-19b and miR-494 suggested that upregulated miR-19b, cooperating with downregulated miRNA-494, could inhibit PTEN expression. In conclusion, the findings suggest that the overexpression of PTEN plays an important role in the pathogenesis of RSA, with miR-19b directly regulating PTEN and working with miR-494, all of which participating in abnormal effects of villous' trophoblastic cell may be a critical event.

中文翻译：

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受 miR-19b 和 miR-494 调控的 PTEN 在复发性自然流产患者绒毛中的过表达。

复发性自然流产（RSA）可归因于多种因素，滋养细胞的异常侵袭和凋亡引起了广泛关注。我们的研究旨在调查具有潜在调节关系的 PTEN 和 miRNA 在 RSA 患者胎盘绒毛中的表达。本研究招募了 19 名 RSA 患者和 16 名育龄期健康妇女进行人工流产 (IA)。使用实时聚合酶链反应 (PCR) 和蛋白质印迹法研究 PTEN 和 miRNA 的表达，进一步验证 PTEN 与使用细胞培养和转染的潜在 miRNA 之间的关系，并使用荧光素酶活性测定来确定 PTEN 是否受潜在的直接调节。 miRNA。结果表明，RSA 患者 PTEN mRNA 和蛋白质表达水平均上调，但仅在蛋白质表达水平上观察到显着差异（p < 0.001）。通过实时 PCR 预扫描，9 种潜在 miRNA 的结果揭示了 3 种显着上调的 miRNA（miR-494、miR-146a 和 miR-21）和一种显着下调的 miRNA（miR-19b）。关于miR-19b和miR-494的进一步验证结果表明，上调的miR-19b与下调的miRNA-494配合，可以抑制PTEN的表达。综上所述，研究结果表明PTEN的过表达在RSA的发病机制中起重要作用，miR-19b直接调节PTEN并与miR-494共同参与绒毛病的异常作用。