Transcriptomic profiling of trophoblast fusion using BeWo and JEG-3 cell lines. (29th November 2019)

• Record Type: Journal Article
• Authors: Msheik, H; El Hayek, S; Bari, M Furqan; Azar, J; Abou-Kheir, W; Kobeissy, F; Vatish, M; Daoud, G
• Is Part Of: Molecular human reproduction. Volume 25:Number 12(2019)
• Page Start: 811
• Page End: 824
• Publication Date: 2019-11-29
• Subjects: BeWo -- JEG-3 -- trophoblast -- SIK1 -- gene expression -- fusion -- human chorionic gonadotropin -- microarrays; Human reproduction -- Molecular aspects -- Periodicals; Electronic journals; 612.6
• DOI: 10.1093/molehr/gaz061
• Languages: English
• ISSNs: 1360-9947

Abstract

Abstract: In human placenta, alteration in trophoblast differentiation has a major impact on placental maintenance and integrity. However, little is known about the mechanisms that control cytotrophoblast fusion. The BeWo cell line is used to study placental function, since it forms syncytium and secretes hormones after treatment with cAMP or forskolin. In contrast, the JEG-3 cell line fails to undergo substantial fusion. Therefore, BeWo and JEG-3 cells were used to identify a set of genes responsible for trophoblast fusion. Cells were treated with forskolin for 48 h to induce fusion. RNA was extracted, hybridised to Affymetrix HuGene ST1.0 arrays and analysed using system biology. Trophoblast differentiation was evaluated by real-time PCR and immunocytochemistry analysis. Moreover, some of the identified genes were validated by real-time PCR and their functional capacity was demonstrated by western blot using phospho-specific antibodies and CRISPR/cas9 knockdown experiments. Our results identified a list of 32 altered genes in fused BeWo cells compared to JEG-3 cells after forskolin treatment. Among these genes, four were validated by RT-PCR, including salt-inducible kinase 1 (SIK1) gene which is specifically upregulated in BeWo cells upon fusion and activated after 2 min with forskolin. Moreover, silencing of SIK1 completely abolished the fusion. Finally, SIK1 was shown to be at the center of many biological and functional processes, suggesting that it might play a role in trophoblast differentiation. In conclusion, this study identified new target genes implicated in trophoblast fusion. More studies are required to investigate the role of these genes in some placental pathology.