The effect of IL-6 on the trophoblast cell line HTR-8/SVneo

Embryonic development up to the blastocyst stage, implantation into the uterine wall and the development of the functional placenta are steps crucial for the establishment of normal pregnancy. Specific cells of the placenta, the trophoblast cells, invade the uterine stroma and spiral arteries, adapting them to pregnancy. Interleukin-6 is present in the human endometrium during the receptive phase and early pregnancy. Trophoblasts also produce IL-6, which was found to stimulate trophoblast invasion and migration in vitro. Here we show that the activity of MMP-9 may contribute to the observed increased invasion. In addition, in the HTR-8/SVneo trophoblast cell line IL-6 increases cell proliferation.

The effect of prolactin on human trophoblast in vitro

Prolaktin (PRL) je polipeptidni hormon koji utiče na rast i diferencijaciju različitih ćelija, a uključen je i u brojne fiziološke procese. Sintetiše ga i luči hipofiza, ali i različita druga tkiva i ćelije. Prolaktin je jedan od glavnih proteina koji sintetiše i sekretuje decidualizovani endometrijum. Produkcija decidualnog prolaktina se povećava u trudnoći nakon implantacije, i dostiže maksimum u tkivu decidue 20 do 25 nedelje gestacije. Delovanje PRL se ostvaruje vezivanjem za PRL receptore (PRLR) na ćelijskoj membrani. Poznato je da se za prolaktinski receptor vezuje najmanje tri liganda - prolaktin, placentni laktogen i hormon rasta. Do sada je za placentu poznato da je receptor za prolaktin ispoljen u decidui, trofoblastu horiona, epitelu amniona i sinciciotrofoblastu na kraju trudnoće. Prolaktin u različitim ćelijama i tkivima utiče na ekspresiju adhezionih i proteolitičkih molekula koji su značajni za degradaciju ekstraćelijskog matriksa i ćelijsku migraciju. Međutim, njegova uloga u trofoblastu nije dovoljno poznata. Ovim radom je po prvi put ispitivan uticaj prolaktina na svojstva invazivnog ekstravilusnog trofoblasta prvog trimestra trudnoće. Detektovana je ekspresija PRLR, kao i profil prisutnih formi receptora na izolovanim citotrofoblastnim ćelijama i HTR-8/SVneo trofoblastnoj imortalizovanoj ćelijskoj liniji. Pokazali smo da PRL stimuliše adheziju, migraciju i invaziju trofoblasta in vitro. Kao potencijalne efektore ispitivali smo integrine, gal-1 i matriksne metaloproteinaze-2 i -9. Dobijeni rezultati ukazuju da PRL stimuliše ekspresiju subjedinica integrina α1, α5, kao i gal-1. Ispitivali smo i efekat PRL na vijabilnost, proliferaciju i apoptozu HTR-8/SVneo ćelija. PRL je blago stimulisao vijabilnost i uticao na povećanje broja adherentnih ćelija, dok apoptoza nije bila značajno promenjena. Dobijeni rezultati predstavljaju napredak u razumevanju fiziološke uloge PRL značajne za funkciju humanog trofoblasta. Takođe, ovaj rad pruža celovitiji uvid u proces diferencijacije ranog trofoblasta i predstavlja dobru osnovu za dalja ispitivanja biološke uloge prolaktina, kao i drugih članova ove familije proteina u humanom trofoblastu.Prolactin (PRL) is a polypeptide hormone which has impact on the growth and differentiation of various cell types, and is known to participate in numerous physiological processes. It is synthesized and secreted by the pituitary gland and many other tissues and cell types. Prolactin is also one of the major proteins synthesized and secreted by decidualized endometrium. The production of decidual prolactin increases after implantation, and peaks in decidual tissue at 20 to 25th week of gestation. The action of PRL is exerted through binding to its receptors (PRLR) at the surface of target cells. It is known that prolactin receptor has at least three ligands – prolactin, placental lactogen and growth hormone. Regarding localization of prolactin receptor in placenta, so far it has been found in decidua, chorionic trophoblast, amniotic epithelium and synciciotrophoblast at term. Prolactin affects the expression of adhesion and proteolytic molecules which are important for degradation of extracellular matrix and cell migration. However, its role in trophoblast is not well defined. In this study the effect of prolactin on the function of first trimester of pregnancy extravillous trophoblast was studied for the first time. The expression of PRLR, as well as the profile of different isoforms was examined in both cytotrophoblast and HTR-8/SVneo trophoblast derived cell line. It is shown here that PRL stimulates trophoblast cell adhesion, migration and invasion in vitro. Potential effectors were sought among integrins, gal-1 and matrix metalloproteinases-2 and -9. The results showed that PRL stimulated the expression of integrin subunits α1, α5, as well as gal-1. When investigating the effect of PRL on cell viability, proliferation and apoptosis of HTR-8/SVneo cells, PRL was found to slightly stimulate cell viability and adherent cell number, while apoptosis was not altered. The results of this study represent a step further in the understanding of the physiological role of PRL in human trophoblast. Moreover, this work gives a better insight in to the process of early trophoblast differentiation and represents a good basis for further studies of the biological role of prolactin, as well as other members of this family of proteins in the human trophoblast

Galectin-1 Is Part of Human Trophoblast Invasion Machinery - A Functional Study In Vitro

Interactions of glycoconjugates with endogenous galectins, have been long proposed to participate in several reproductive processes including implantation. In human placenta gal-1, gal-3, gal-8, and gal-13 proteins are known to be present. Each of them has been proposed to play multiple functions, but so far no clear picture has emerged. We hypothesized that gal-1 participates in trophoblast invasion, and conducted Matrigel invasion assay using isolated cytotrophoblast from first trimester placenta and HTR-8/SVneo cell line to test it.<0.001) by Ox-gal-1 at 1 µg/ml. Both sets of results confirmed involvement of gal-1 in trophoblast invasion. Galectin profile of isolated cytotrophoblast and HTR-8/SVneo cells was established using RT-PCR and real-time PCR and found to consist of gal-1, gal-3 and gal-8 for both cell types. Only gal-1 was located at the trophoblast cell membrane, as determined by FACS analysis, which is consistent with the results of the functional tests.These findings qualify gal-1 as a member of human trophoblast cell invasion machinery

Expression of prolactin receptors in the human extra villous cytotrophoblast cell line HTR-8/SVNEO

Prolactin is a pituitary hormone that is widely produced at extrapitutitary locations. One of the first described sites of its production is the decidualized endometrium, suggesting that it could be potentially significant for local processes such as immunomodulation, embryo implantation, and survival. In this study, expression of prolactin receptor protein is investigated by immunocytochemistry and Western blot in the HTR-8/SVneo cell line, often used as a model of first-tri­mester trophoblast. The obtained data enabled us to identify both long and intermediate forms of prolactin receptors, which were shown to be significant in PRL signaling in other tissues

Expression of prolactin receptors in the human extravillous cytotrophoblast cell line HTR-8/SVneo

Abstract — Prolactin is a pituitary hormone that is widely produced at extrapitutitary locations. One of the first described sites of its production is the decidualized endometrium, suggesting that it could be potentially significant for local processes such as immunomodulation, embryo implantation, and survival. In this study, expression of prolactin receptor protein is investigated by immunocytochemistry and Western blot in the HTR-8/SVneo cell line, often used as a model of first-trimester trophoblast.The obtained data enabled us to identify both long and intermediate forms of prolactin receptors, which were shown to be significant in PRL signaling in other tissues

DOI:10.2298/ABS1003531J THE EFFECT OF IL-6 ON THE TROPHOBLAST CELL LINE HTR-8/SVNEO

Abstract- Embryonic development up to the blastocyst stage, implantation into the uterine wall and the development of the functional placenta are steps crucial for the establishment of normal pregnancy. Specific cells of the placenta, the trophoblast cells, invade the uterine stroma and spiral arteries, adapting them to pregnancy. Interleukin-6 is present in the human endometrium during the receptive phase and early pregnancy. Trophoblasts also produce IL-6, which was found to stimulate trophoblast invasion and migration in vitro. Here we show that the activity of MMP-9 may contribute to the observed increased invasion. In addition, in the HTR-8/SVneo trophoblast cell line IL-6 increases cell proliferation

The edible mushroom Laetiporus sulphureus as potential source of natural antioxidants

Hot water extract (LN), partially purified polysaccharides (LP) and hot alkali extracted polysaccharides (LNa) obtained from fruiting bodies of the wild basidiomycete Laetiporus sulphureus were examined for their antioxidant activities. LNa was the most active antioxidant, as shown by the median effective concentrations (EC50 values) of 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity (0.5 +/- 0.2 mg/ml), reducing power (4.0 +/- 0.3 mg/ml) and ferrous ion-chelating ability (1.5 +/- 0.1 mg/ml). LNa contained the highest level of alpha-glucan (17.3 +/- 1.2 g/100 g dw), whereas LP contained mostly beta-glucans (66.8 +/- 1.3 g/100 g dw). The prevalent monosaccharide in all extracts was glucose. The EC50 values of all three antioxidant activity assays were well-correlated with the alpha-glucan content. Strong and significant correlation was found between total phenolic compounds and DPPH scavenging ability and also reducing power. The three investigated extracts (at concentrations of 0.1-10 mg/ml) were not toxic to HTR-8/SVneo trophoblast cell line

MIF is among the proinflammatory cytokines increased by LPS in the human trophoblast line

Infection is increasingly considered to contribute to pathological conditions in pregnancy. The placenta acts as a protective immunological fetomaternal barrier which recognizes microbes by pattern recognition receptors on the trophoblast. Lipopolysaccharide (LPS) is a cell wall constituent of Gram-negative bacteria that elicits a strong immune response. In this study, LPS from E. coli was used to treat the HTR-8/SVneo trophoblast cell line and examine its influence on cytokines IL-6, IL-8 and MIF using real-time PCR, metalloproteinases (MMP)-2 and -9 by gelatin zymography, and Western analysis of integrin subunits α1 and β1, all known to contribute to migration of human trophoblasts in vitro. The results described herein for the first time, show that MIF mRNA and secreted MIF protein were significantly elevated (2.5-3- and 2-fold, respectively) in LPS-treated cells. MMP-2 and MMP-9 levels were increased, as well as cell migration, as judged by a wound-healing test, however, no changes in the studied integrin subunits, cell viability or cell numbers were observed. The data obtained furthers our understanding of LPS actions on the trophoblast in vitro, additionally implicate MIF, and suggest that infection in vivo could indeed alter the functional characteristics of the trophoblast. [Projekat Ministarstva nauke Republike Srbije, br. 173004