HAI-1 regulates placental folds development by influencing trophoblast cell proliferation and invasion in pigs.

Abstract

Fetal development is critically dependent on the efficiency of the placenta. Porcine trophoblast cell proliferation and invasion have crucial roles in placental fold development, which is one of the essential events determining placental efficiency. The membrane serine proteinase inhibitor hepatocyte growth factor activator inhibitor-1 (HAI-1) can regulate cellular invasion and motility in different types of epithelial cells, including trophoblast cells in mice. This work used quantitative polymerase chain reaction (qPCR) and immunohistochemistry to compare the expression level and location of HAI-1 in the placenta on gestational days 26, 50, and 95 in Yorkshire and Meishan pigs. The role of HAI-1 in porcine trophoblast cell (PTr2) proliferation, invasion, and migration in vitro was investigated by analyzing the effects of HAI-1 gene silencing or overexpression. Polymorphism in the HAI-1gene was detected to determine associations between the genotype and piglet birth weight in 400 healthy pure-bred Yorkshire piglets. qPCR results showed that HAI-1 mRNA levels significantly increased (P<0.01) between gestational days 26 and 50 and then decreased (P<0.01) between days 50 and 95 in both Meishan and Yorkshire pigs. Immunohistochemical analysis showed that HAI-1 protein was strongly expressed by the high columnar trophoblast cells located at the top of the placental folds with low proliferative and invasion capacities. However, it was expressed at very low levels in cuboidal trophoblast cells located at the side and base of the placental folds with high proliferative and invasion capacities. In vitro experiments indicated that HAI-1 had the ability to reduce the proliferation, invasion and migration of trophoblast cells. In addition, one single-nucleotide polymorphism (SNP) of HAI-1 showed a significant association (P<0.05) with piglet birth weight. These results revealed that HAI-1 could be a vital molecule in placental folds development by regulating trophoblast proliferation and invasion in pigs.