RESUMO
Invasive extravillous cytotrophoblast of the human placenta expresses galectins-1, -3, and -8 in vivo and in vitro. This study aimed to investigate the potential role of galectin-3 in cell migration and invasion, using recombinant human galectin-3 (rhgalectin-3), small molecule galectin inhibitor I47, and galectin-3 silencing. HTR-8/SVneo cell migration was stimulated by rhgalectin-3 and reduced by I47, which could be neutralised by rhgalectin-3. Inhibitor specificity and selectivity for the galectins expressed in extravillous trophoblast were validated in solid phase assays using recombinant galectin-1, -3, -8, confirming selectivity for galectin-3. HTR-8/SVneo cell migration and invasion, and invasion by isolated trophoblast cells in primary culture were significantly reduced in the presence of I47, which could be restored by rhgalectin-3. Upon HTR-8/SVneo cell treatment with galectin-3 siRNA both LGALS3 and galectin-3 protein were dramatically decreased. Silencing of galectin-3 induced significant reduction in cell migration and invasion, which was restored by rhgalectin-3. The influence on known mediators of cell invasion, MMP2 and -9, and integrins α1, α5, and ß1 was followed in silenced cells, showing lower levels of MMPs and a large reduction in integrin subunit ß1. These results show that galectin-3 acts as a pro-invasive autocrine/paracrine factor in trophoblast in vitro.
Assuntos
Movimento Celular , Sobrevivência Celular , Galectina 3/metabolismo , Trofoblastos/patologia , Proteínas Sanguíneas , Células Cultivadas , Feminino , Galectina 3/antagonistas & inibidores , Galectina 3/genética , Galectinas , Humanos , Integrinas/metabolismo , Gravidez , RNA Interferente Pequeno/genética , Trofoblastos/metabolismoAssuntos
Terapia Baseada em Transplante de Células e Tecidos/métodos , Fertilização in vitro/métodos , Células Germinativas , Células-Tronco Pluripotentes Induzidas , Infertilidade Feminina/terapia , Infertilidade Masculina/terapia , Animais , Modelos Animais de Doenças , Feminino , Gametogênese/fisiologia , Humanos , Masculino , Casamento , Camundongos , TrissomiaRESUMO
Galectins (gals) as multifunctional animal lectins are of great potential significance for establishment and function of the placenta, due to their capacity to modulate cellular functions including proliferation, adhesion, and invasion. Human trophoblast is known to express gal-1, gal-3 and gal-13 proteins, as well as mRNAs for gal-14, -16 and -17. This study was undertaken to establish cellular distribution of gal-8, not previously associated with trophoblast, since we have recently detected gal-8 RNA and protein in cytotrophoblast cell material. Results of immunolocalization showed that gal-8 was expressed by villous and extravillous cytotrophoblast.
