Dissimilar microRNA-21 functions and targets in trophoblastic cell lines of different origin
Dissimilar microRNA-21 functions and targets in trophoblastic cell lines of different origin
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DOI：
10.1016/j.biocel.2015.08.018
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发表时间：
2015-11-01
影响因子：
4
通讯作者：
Morales-Prieto, Diana M.
中科院分区：
文献类型：
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作者：
Chaiwangyen, Wittaya;Ospina-Prieto, Stephanie;Morales-Prieto, Diana M.
滋养层细胞表达一种单一的 miRNA 表达谱，该表达谱在妊娠期间发生变化，其变化可能与妊娠并发症有关。 miR-21 是一种广为人知的 oncomir，在人胎盘中高表达，但其在调节滋养层细胞中的作用仍不清楚。本研究的目的是研究 miR-21 在 HTR-8/SVneo 永生化滋养层细胞和 JEG-3 绒毛膜癌细胞中的功能和靶点，这两种细胞是细胞起源不同的滋养层细胞模型。用miR-21-antagomir、-mimic或它们各自的对照转染细胞。随后，评估了细胞增殖（BrdU）、迁移（Transwell 和划痕伤口愈合测定）、侵袭（Matrigel 测定）和细胞凋亡（流式细胞术、TUNEL 测定和蛋白质印迹）。通过蛋白质印迹分析潜在的 miR-21 靶点磷酸酶和张力蛋白同源物 (PTEN) 和程序性细胞死亡 4 (PDCD4) 的表达。抑制 miR-21 可减少 JEG-3 和 HTR-8/SVneo 细胞的细胞增殖、迁移和侵袭，此外还诱导 JEG-3 细胞凋亡。沉默 miR-21 仅在 JEG-3 细胞中增强 PDCD4 表达，仅在 HTR-8/SVneo 细胞中增强 PTEN 表达。抑制 miR-21 显着增加 HTR-8/SVneo 细胞中 AKT 的磷酸化。总之，miR-21 具有细胞特异性靶标，具体取决于滋养层细胞的起源。此外，miR-21 调节主要细胞过程，包括细胞生长、迁移、侵袭和凋亡，表明其损伤可能导致胎盘疾病。 (C) 2015 Elsevier Ltd. 保留所有权利。
Trophoblast cells express a singular miRNA expression profile which varies during pregnancy and whose alteration may be associated with pregnancy complications. miR-21, a widely known oncomir, is highly expressed in human placenta but its role in regulating trophoblast cells remains unclear. The aim of this study was to investigate miR-21 functions and targets in HTR-8/SVneo immortalized trophoblast and JEG-3 choriocarcinoma cells, which are trophoblast cell models that differ in their cellular origin. Cells were transfected with miR-21-antagomir, -mimic or their respective controls. Following, cell proliferation (BrdU), migration (Transwell and scratch wound-healing assays), invasion (Matrigel assays) and apoptosis (flow cytometry, TUNEL assay and Western blotting) were assessed. Expression of the potential miR-21 targets phosphatase and tensin homolog (PTEN) and programmed cell death 4 (PDCD4) were analyzed by Western blotting. Inhibition of miR-21 decreased cell proliferation, migration, and invasion in JEG-3 and HTR-8/SVneo cells and additionally, induced apoptosis in JEG-3 cells. Silencing of miR-21 enhanced PDCD4 expression only in JEG-3 cells, and PTEN expression only in HTR-8/SVneo cells. Inhibition of miR-21 significantly increased phosphorylation of AKT in HTR-8/SVneo cells. In conclusion, miR-21 has cell-specific targets depending upon the origin of trophoblastic cells. Furthermore, miR-21 regulates major cellular processes including cell growth, migration, invasion and apoptosis suggesting that its impairment may lead to placental disorders. (C) 2015 Elsevier Ltd. All rights reserved.