Activin A promotes human trophoblast invasion by upregulating integrin β3 via ALK4-SMAD4 signaling

Activin A has been widely regarded as an important promoter of trophoblast invasion during the first trimester of pregnancy. However, whether integrin β3 is involved in activin A-upregulated trophoblast invasion and the underlying molecular mechanisms remain largely unknown. We utilized immortalized (HTR8/SVneo) and primary human extravillous trophoblast (EVT) cells, as well as first-trimester chorionic villous explants as study models to investigate the function and underlying molecular mechanisms of integrin β3 in activin A-promoted human trophoblast invasion. We found that activin A increased integrin β3 mRNA and protein levels in both HTR8/SVneo and primary EVT cells, and knockdown of integrin β3 significantly decreased basal and activin A-upregulated trophoblast invasion. Moreover, SB431542 (a specific inhibitor of TGF-β type Ι receptor kinase) abolished activin A-upregulated integrin β3 expression and SMAD2/3 phosphorylation. In addition, siRNA-mediated knockdown of ALK4 or SMAD4 both abolished activin A-upregulated integrin β3 expression in HTR8/SVneo cells, while knockdown of ALK4 or SMAD4 attenuated activin A-upregulated integrin β3 expression in primary EVTs. Our findings reveal the mediation role of integrin β3 in activin A-upregulated human trophoblast invasion and that activin An upregulates integrin β3 expression in an ALK4-SMAD4 signaling-dependent manner.