Wang Ying, Zhang Yang, Wang Huimin, Wang Junxiao, Zhang Yiyuan, Wang Yingzhe, Pan Zhenwei, Luo Shanshun
2. Department of Pharmacology (Key Laboratory of Cardiovascular Medicine Research, Ministry of Education; State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, Heilongjiang 150081, P. R. China.
1. Department of Gerontology, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150081, P. R. China;
Int J Biol Sci. 2014 Aug 30;10(9):973-82. doi: 10.7150/ijbs.9088. eCollection 2014.
Preeclampsia is a serious complication in pregnancy. Dysregulation of trophoblast cell proliferation and invasion is a major pathological alteration observed in preeclampsia. Recently, microRNAs were shown to participate in the pathogenesis of preeclampsia. In this study we explored the effect of miR-20a on the proliferation and invasion of trophoblast cells and the underlying mechanism. We verified the distribution of miR-20a in human placenta by in situ hybridization. Real time PCR data showed that the level of miR-20a increased by 2.6 folds in human preeclampsia than normal tissues. We then cultured trophoblast-like JEG-3 cells and evaluated the effect of miR-20a on JEG-3 cell proliferation, migration and invasion. Overexpression of miR-20a significantly inhibited the proliferation, migration and invasion of cultured JEG-3 cells, which were abolished by co-transfection of AMO-20a. Transfection of miR-20a also inhibited JEG-3 cell xenograft tumor growth in nude mice. Luciferase assay technique was used to identify the direct regulation of miR-20a on Forkhead Box Protein A1(FOXA1). Transfection of miR-20a markedly reduced the luciferase activity of the chimeric plasmid containing the 3'UTR of FOXA1, indicating FOXA1 is the target of miR-20a. Furthermore, transfection of miR-20a inhibited both protein and mRNA expression of FOXA1 in JEG-3 cells. In summary, the upregulated miR-20a in human preeclampsia tissue can inhibit the proliferative and invasive activities of trophoblast cells by repressing the expression of FOXA1.
子痫前期是妊娠期一种严重的并发症。滋养层细胞增殖和侵袭失调是子痫前期观察到的主要病理改变。最近，微小RNA被证明参与子痫前期的发病机制。在本研究中，我们探讨了miR-20a对滋养层细胞增殖和侵袭的影响及其潜在机制。我们通过原位杂交验证了miR-20a在人胎盘组织中的分布。实时PCR数据显示，子痫前期患者组织中miR-20a的水平比正常组织增加了2.6倍。然后我们培养了滋养层样JEG-3细胞，并评估了miR-20a对JEG-3细胞增殖、迁移和侵袭的影响。miR-20a的过表达显著抑制了培养的JEG-3细胞的增殖、迁移和侵袭，而共转染AMO-20a可消除这些作用。转染miR-20a也抑制了JEG-3细胞在裸鼠体内异种移植瘤的生长。采用荧光素酶报告基因检测技术鉴定miR-20a对叉头框蛋白A1（FOXA1）的直接调控作用。转染miR-20a显著降低了含有FOXA1 3'UTR的嵌合质粒的荧光素酶活性，表明FOXA1是miR-20a的靶标。此外，转染miR-20a抑制了JEG-3细胞中FOXA1的蛋白和mRNA表达。总之，子痫前期患者组织中上调的miR-20a可通过抑制FOXA1的表达来抑制滋养层细胞的增殖和侵袭活性。
