目的:滋养细胞功能紊乱是导致子痫前期(PE)的重要因素之一。已发现细胞质聚腺苷酸化元件结合2(CPEB2)在PE患者中差异表达,但它是否通过调节滋养细胞功能介导PE过程尚不清楚。
方法:采用实时定量PCR检测CPEB2和生长抑素受体3(SSTR3)的表达,Westernblot(WB)和免疫荧光染色。通过CCK-8测定分析细胞功能,EdU分析,流式细胞术和transwell分析。通过WB检测上皮-间质转化(EMT)相关蛋白水平。CPEB2和SSTR3的相互作用通过RIP实验证实,双荧光素酶报告基因测定和PCRpoly(A)尾测定。进行动物实验以探索CPEB2对体内PE进展的影响。大鼠胎盘组织进行H&E染色,免疫组化染色和TUNEL染色。
结果:CPEB2在PE患者中低表达。CPEB2上调加速滋养细胞增殖,迁移,入侵和EMT,而它的击倒却产生了相反的效果。CPEB2与SSTR3mRNA的3'-UTR中的CPE位点结合,通过减少poly(A)尾巴来抑制SSTR3的翻译。此外,SSTR3过表达抑制滋养层细胞增殖,迁移,入侵和EMT,而它的沉默加速了滋养层细胞的功能。然而,这些影响可以通过CPEB2的上调和敲低来逆转,分别。体内实验,CPEB2过表达减轻组织病理学改变,抑制细胞凋亡,通过降低SSTR3的表达促进PE大鼠胎盘的增殖并增强EMT。
结论:CPEB2抑制PE进展,通过多腺苷酸化抑制SSTR3翻译促进滋养层细胞功能。
Trophoblast cell dysfunction is one of the important factors leading to preeclampsia (PE). Cytoplasmic polyadenylation element-binding 2 (
CPEB2) has been found to be differentially expressed in PE patients, but whether it mediates PE process by regulating trophoblast cell function is unclear.
The expression of
CPEB2 and somatostatin receptor 3 (SSTR3) was detected by quantitative real-time PCR, Western blot (WB) and immunofluorescence staining. Cell functions were analyzed by CCK-8 assay, EdU assay, flow cytometry and transwell assay. Epithelial-mesenchymal transition (EMT)-related protein levels were detected by WB. The interaction of
CPEB2 and SSTR3 was confirmed by RIP assay, dual-luciferase reporter assay and PCR poly(A) tail assay. Animal experiments were performed to explore the effect of CPEB2 on PE progression in vivo, and the placental tissues of rat were used for H&E staining, immunohistochemical staining and TUNEL staining.
CPEB2 was lowly expressed in PE patients. CPEB2 upregulation accelerated trophoblast cell proliferation, migration, invasion and EMT, while its knockdown had an opposite effect. CPEB2 bound to the CPE site in the 3\'-UTR of SSTR3 mRNA to suppress SSTR3 translation through reducing poly(A) tails. Besides, SSTR3 overexpression suppressed trophoblast cell proliferation, migration, invasion and EMT, while its silencing accelerated trophoblast cell functions. However, these effects could be reversed by
CPEB2 upregulation and knockdown, respectively. In vivo experiments, CPEB2 overexpression relieved histopathologic changes, inhibited apoptosis, promoted proliferation and enhanced EMT in the placenta of PE rat by decreasing SSTR3 expression.
CPEB2 inhibited PE progression, which promoted trophoblast cell functions by inhibiting SSTR3 translation through polyadenylation.