Knockdown of activated Cdc42-associated kinase inhibits human extravillous trophoblast migration and invasion and decreases protein expression of pho-Akt and matrix metalloproteinase

Hsa-miR-210-3p has been reported to be upregulated in preeclampsia (PE); however, the functions of miR-210-3p in placental development are not fully understood, and, consequently, miR-210-3p’s role in the pathogenesis of PE is still under investigation. In this study, we found that overexpression of miR-210-3p reduced trophoblast migration and invasion, extravillous trophoblast (EVT) outgrowth in first trimester explants, expression of endovascular trophoblast (enEVT) markers and the ability of trophoblast to form endothelial-like networks. In addition, miR-210-3p overexpression significantly downregulated the mRNA levels of interleukin-1B and -8, as well as CXC motif ligand 1. These cytokines have been suggested to play a role in EVT invasion and the recruitment of immune cells to the spiral artery remodeling sites. We also showed that caudal-related homeobox transcription factor 2 (CDX2) is targeted by miR-210-3p and that CDX2 downregulation mimicked the observed effects of miR-210-3p upregulation in trophoblasts. These findings suggest that miR-210-3p may play a role in regulating events associated with enEVT functions and its overexpression could impair spiral artery remodeling, thereby contributing to PE.

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Extravillous Trophoblast Migration and Invasion Assay

BIO-PROTOCOL ◽

10.21769/bioprotoc.840 ◽

2013 ◽

Vol 3 (15) ◽

Cited By ~ 3

Author(s):

Magdalena Angelova ◽

Heather Machado ◽

Kenneth Swan ◽

Cindy Morris ◽

Deborah Sullivan

Keyword(s):

Extravillous Trophoblast ◽

Migration And Invasion ◽

Invasion Assay ◽

Trophoblast Migration

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Spatiotemporal expression profile of proteases and immunological, angiogenic, hormonal and apoptotic mediators in rat placenta before and during intrauterine trophoblast migration

Reproduction Fertility and Development ◽

10.1071/rd16280 ◽

2017 ◽

Vol 29 (9) ◽

pp. 1774 ◽

Cited By ~ 2

Author(s):

Juneo F. Silva ◽

Natália M. Ocarino ◽

Rogéria Serakides

Keyword(s):

Gene Expression ◽

Nitric Oxide ◽

Growth Factor ◽

Nitric Oxide Synthase ◽

Matrix Metalloproteinase ◽

Protein Expression ◽

Trophoblast Invasion ◽

Toll Like Receptor ◽

Oxide Synthase ◽

Trophoblast Migration

The gene and/or protein expression of proteases and immunological, angiogenic, hormonal and apoptotic mediators was evaluated in rat placenta before and during intrauterine trophoblast migration. The depth of interstitial and endovascular intrauterine trophoblast invasion and the immunohistochemical expression of vascular endothelial growth factor (VEGF), fetal liver kinase 1 (Flk1), interferon (IFN)-γ, migration inhibitory factor (MIF), and inducible nitric oxide synthase (iNOS; also known as nitric oxide synthase (NOS) 2) were evaluated. In addition, the expression of the Vegf, Flk1, placental growth factor (Pigf), soluble fms-like tyrosine kinase 1 (sFlt1), placental lactogen 1 (Pl1), proliferin-related protein (rPlf), placental leptin (Lep), Toll-like receptor 2 (Tlr2), Toll-like receptor 4 (Tlr4), Infg, Mif, tumour necrosis factor-α (Tnf), interleukin-10 (Il10), Nos2, caspase 3 (Casp3), Bax, Bcl2, matrix metalloproteinase 2 (Mmp2) and matrix metalloproteinase 9 (Mmp9) genes was determined by real-time reverse transcription–polymerase chain reaction. At 10 days gestation, gene expression of Tlr2, Tlr4, Tnf, Infg, Il10, Casp3, Pigf, sFlt1 and Lep (P < 0.05) were higher than at 14 and/or 19 days of gestation. The beginning of intrauterine trophoblast invasion, i.e., at 14 days of gestation, coincided with higher gene and/or protein expression of MMP9, VEGF, Flk1, NOS2, MIF, BAX and rPlf compared to days 10 and 19 (P < 0.05). In contrast, gene expression of Mmp2 and Pl1 was higher at the end of trophoblast invasion compared to 10 and 14 days of gestation (P < 0.05). In conclusion, before intrauterine trophoblast migration, expression of TLRs and immunological and pro-apoptotic mediators is higher, whereas the beginning of trophoblast migration is characterised by higher expression of the pro-angiogenic factors NOS2 and MMP9. In contrast, MMP2 and PL1 expression is higher at the end of intrauterine trophoblast migration.

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CRY2 suppresses trophoblast migration and invasion in recurrent spontaneous abortion

The Journal of Biochemistry ◽

10.1093/jb/mvz076 ◽

2019 ◽

Vol 167 (1) ◽

pp. 79-87 ◽

Cited By ~ 2

Author(s):

Lianzhi Wu ◽

Biheng Cheng ◽

Qian Liu ◽

Ping Jiang ◽

Jing Yang

Keyword(s):

Spontaneous Abortion ◽

Cell Model ◽

Recurrent Spontaneous Abortion ◽

Extravillous Trophoblast ◽

Matrix Metalloproteinase 2 ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Protein Levels ◽

Cryptochrome 2 ◽

Trophoblast Migration

Abstract Disruption of circadian rhythms is associated with aberrant trophoblast migration and invasion in recurrent spontaneous abortion (RSA). This study aims to explore the functional role and the mechanisms of cryptochrome 2 (CRY2), a fundamental component of the circadian clock, in regulating trophoblast migration and invasion. Human extravillous trophoblast cell line HTR-8/SVneo was used as a cell model. Cell migration and invasion were examined using wound healing assay and Transwell assay, respectively. The mRNA and protein levels were determined using quantitative real-time polymerase chain reaction and western blot, respectively. Luciferase reporter assay and chromatin immunoprecipitation assay were performed to explore the interaction between c-Myc to the brain and muscle ARNT-like protein 1 (BMAL1) promoter. CRY2 was highly expressed in human villous specimens of RSA. Furthermore, CRY2 overexpression impaired migration and invasion in HTR-8/SVneo cells, whereas CRY2 knockdown yielded the opposite results. Mechanistically, c-Myc bound to the BMAL1 promoter and induced BMAL1 transcription, both of which further activated matrix metalloproteinase 2/9 (MMP2/9) and facilitated migration and invasion in HTR-8/SVneo cells. CRY2 inhibited c-Myc-BMAL1 pathway and impaired migration and invasion of HTR-8/SVneo cells. Collectively, these findings demonstrate that CRY2 suppresses trophoblast migration and invasion via inhibiting c-Myc-BMAL1-MMP2/9 pathway.

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Extravillous Trophoblast Migration and Invasion: Impact of Environmental Chemicals and Pharmaceuticals

Reproductive Toxicology ◽

10.1016/j.reprotox.2021.11.008 ◽

2021 ◽

Author(s):

Cassandra Meakin ◽

Emily S. Barrett ◽

Lauren M. Aleksunes

Keyword(s):

Environmental Chemicals ◽

Extravillous Trophoblast ◽

Migration And Invasion ◽

Invasion Impact ◽

Trophoblast Migration

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Regulation of the Matricellular Proteins CYR61 (CCN1) and NOV (CCN3) by Hypoxia-Inducible Factor-1α and Transforming-Growth Factor-β3 in the Human Trophoblast

Endocrinology ◽

10.1210/en.2009-1195 ◽

2010 ◽

Vol 151 (6) ◽

pp. 2835-2845 ◽

Cited By ~ 36

Author(s):

Nadine Wolf ◽

Wei Yang ◽

Caroline E. Dunk ◽

Isabella Gashaw ◽

Stephen J. Lye ◽

...

Keyword(s):

Hypoxia Inducible Factor ◽

Extravillous Trophoblast ◽

Migration And Invasion ◽

Ccn Proteins ◽

Low Oxygen ◽

Trophoblast Cells ◽

Human Trophoblast ◽

Placental Perfusion ◽

Hypoxia Inducible Factor 1Α ◽

Trophoblast Migration

It is known that a hypoxic environment is critical for trophoblast migration and invasion and is fundamental for appropriate placental perfusion. Because cysteine-rich 61 (CYR61, CCN1) and nephroblastoma overexpressed (NOV, CCN3) are expressed in the extravillous trophoblast and expression levels are deregulated in preeclampsia, we investigated their regulation properties in first-trimester placental explants and in JEG3 choriocarcinoma cells upon a physiological low oxygen tension of 1–3%. In placental explants, both proteins were expressed in the extravillous trophoblast cells and were increased upon hypoxia. JEG3 cells revealed a significant up-regulation of CYR61 and NOV intracellular as well as secreted protein upon hypoxic treatment accompanied by the stabilization of the hypoxia-inducible factor-1α (HIF-1α). Treatment with dimethyloxalylglycine to mimic hypoxia and silencing of HIF-1α using small interfering RNA revealed that only the increase in intracellular protein expression seems to be dependent on HIF-1α but obviously not the secretion process. Moreover, recombinant TGF-β3 was able to further enhance the amount of intracellular CCN proteins as well as secreted CYR61 levels under hypoxia. These results indicate that low oxygen levels trigger elevation of intracellular as well as secreted CYR61 and NOV protein probably in two independent pathways. Addition of recombinant CYR61 and NOV proteins increases migration as well as invasion properties of JEG3 trophoblast cells, which strengthen their role in supporting trophoblast migration invasion properties. In summary, CYR61 and NOV are regulated by HIF-1α and TGF-β3 in the trophoblast cell line JEG3, and their enhanced secretion could be implicated in appropriate placental invasion.

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Inhibition of Migration and Invasion of Hepatocarcinoma HepG2 Cells by Dietary Saponins via Targeting HIF-1α

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520618666180110141827 ◽

2018 ◽

Vol 18 (7) ◽

pp. 1025-1031

Author(s):

Cheng Luo ◽

Di Wu ◽

Meiling Chen ◽

Wenhua Miao ◽

Changfeng Xue ◽

...

Keyword(s):

Cell Proliferation ◽

Confocal Microscopy ◽

Protein Expression ◽

Mtt Assay ◽

Hepg2 Cells ◽

Molecular Mechanisms ◽

Migration And Invasion ◽

Rt Pcr ◽

Gene And Protein Expression ◽

Simulated Hypoxia

Background: Different saponins from herbs have been used as tonic or functional foods, and for treatment of various diseases including cancers. Although clinical data has supported the function of these saponins, their underlying molecular mechanisms have not been well defined. Methods: With the simulated hypoxia created by 8 hours of Cu++ exposure and following 24 hour incubation with different concentration of saponins in HepG2 cells for MTT assay, migration and invasion assays, and for RT-PCR, and with each group of cells for immunofluorescence observation by confocal microscopy. Results： ZC-4 had the highest rate of inhibition of cell proliferation by MTT assay, and the highest inhibition of migration rate by in vitro scratch assay, while ZC-3 had the highest inhibition of invasion ratio by transwell assay. Under the same simulated hypoxia, the molecular mechanism of saponin function was conducted by measuring the gene expression of Hypoxia Inducible Factor (HIF)-1α through RT-PCR, in which ZC-3 showed a potent inhibition of gene HIF-1α. For the protein expression by immunofluorescence staining with confocal microscopy, HIF-1α was also inhibited by saponins, with the most potent one being ZC-4 after eight hours’ relatively hypoxia incubation. Conclusion: Saponins ZC-4 and ZC-3 have the potential to reduce HepG2 cell proliferation, migration and invasion caused by hypoxia through effectively inhibiting the gene and protein expression of HIF-1α directly and as antioxidant indirectly

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Role of ARID1A in the Regulation of Human Trophoblast Migration and Invasion

Reproductive Sciences ◽

10.1007/s43032-021-00686-0 ◽

2021 ◽

Author(s):

Meiyuan Jin ◽

Shouying Xu ◽

Jiayong Li ◽

Lu Li ◽

Chao Tang

Keyword(s):

Migration And Invasion ◽

Human Trophoblast ◽

Trophoblast Migration

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