19 research outputs found
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
Macrophage migration inhibitory factor is involved in endovascular trophoblast cell function in vitro
Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine abundantly present at the feto-maternal interface proposed to play a role in establishment of pregnancy. We have previously shown that pharmacological inhibition of enzymatic activity of MIF decreases extravillous trophoblast invasion and migration in vitro. This study aimed to further elucidate potential role of endogenous trophoblast MIF, and to assess its importance for endovascular trophoblast cell function in particular. Attenuation of MIF by siRNA reduced HTR-8/SVneo cell invasion through Matrigel (59 % of control), expression of integrin α1 (86 % of control) and levels of MMP2 and MMP9 (87 % and 57 % of control, respectively). MIF specific siRNA reduced the ability of HTR-8/SVneo to differentiate in to endothelial-like phenotype, as determined by Matrigel tube formation assay. The total tube length was decreased to 68.6 %, while the number of branching points was reduced to 57.8 % of control. HTR-8/SVneo cell capacity to integrate into HUVEC monolayers was reduced by knock-down of MIF. This could be partly caused by reduced N-cadherin expression to 63 % of control, which decreased with knock-down of MIF, as the expression of this protein was recently shown essential for trophoblast-endothelial interaction. These novel findings indicate a novel role for trophoblast MIF in spiral artery remodeling process
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