Role of LIN28A in Mouse and Human Trophoblast Cell Differentiation1

滋养层 细胞生物学 生物 胚胎干细胞 细胞分化 基因敲除 干细胞 胎盘 免疫学 男科 细胞培养 胎儿 生物化学 遗传学 医学 怀孕 基因
作者
Jill L. Seabrook,Jeremy D. Cantlon,Austin J. Cooney,Erin E. McWhorter,Brittany A. Fromme,Gerrit J. Bouma,Russell V. Anthony,Quinton A. Winger
出处
期刊:Biology of Reproduction [Oxford University Press]
卷期号:89 (4) 被引量:31
标识
DOI:10.1095/biolreprod.113.109868
摘要

Proper regulation of trophoblast proliferation, differentiation, and function are critical for placenta development and function. The RNA-binding protein, LIN28A, has been well characterized as a potent regulator of differentiation in embryonic stem cells; however, little is known about the function of LIN28A in the placenta. We assessed LIN28A in vitro using mouse trophoblast stem (mTS) cells and human trophoblast cells (ACH-3P). We observed that LIN28A decreased and let-7 miRNA increased when mTS cells were induced to differentiate into mouse trophoblast giant cells (mTGCs) upon the removal of FGF4, heparin and conditioned medium. Similarly, we observed that LIN28A decreased in ACH-3P cells induced to syncytialize with forskolin treatment. To assess LIN28A in vivo we examined Embryonic Day 11.5 mouse placenta and observed abundant LIN28A in the chorioallantoic interface and labyrinth layer, with little LIN28A staining in spongiotrophoblast or differentiated mTGCs. Additionally, shRNA-mediated LIN28A knockdown in ACH-3P cells resulted in increased spontaneous syncytialization, and increased levels of syncytiotrophoblast markers hCG, LGALS13, and ERVW-1 mRNA. Additionally, targeted degradation of LIN28A mRNA increased responsiveness to forskolin-induced differentiation. In contrast, targeted degradation of Lin28a mRNA in mTS cells did not alter cell phenotype when maintained under proliferative culture conditions. Together, these data establish that LIN28A has a functional role in regulating trophoblast differentiation and function, and that loss of LIN28A in human trophoblast is sufficient to induce differentiation, but does not induce differentiation in the mouse.

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