TNF-α Regulated Endometrial Stroma Secretome Promotes Trophoblast Invasion

Successful implantation requires the coordinated migration and invasion of trophoblast cells from out of the blastocyst and into the endometrium. This process relies on signals produced by cells in the maternal endometrium. However, the relative contribution of stroma cells remains unclear. The study of human implantation has major technical limitations, therefore the need of in vitro models to elucidate the molecular mechanisms. Using a recently described 3D in vitro models we evaluated the interaction between trophoblasts and human endometrial stroma cells (hESC), we assessed the process of trophoblast migration and invasion in the presence of stroma derived factors. We demonstrate that hESC promotes trophoblast invasion through the generation of an inflammatory environment modulated by TNF-α. We also show the role of stromal derived IL-17 as a promoter of trophoblast migration through the induction of essential genes that confer invasive capacity to cells of the trophectoderm. In conclusion, we describe the characterization of a cellular inflammatory network that may be important for blastocyst implantation. Our findings provide a new insight into the complexity of the implantation process and reveal the importance of inflammation for embryo implantation.

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Per- and Polyfluoroalkyl Substances Differentially Inhibit Placental Trophoblast Migration and Invasion In Vitro

Toxicological Sciences ◽

10.1093/toxsci/kfaa043 ◽

2020 ◽

Vol 175 (2) ◽

pp. 210-219 ◽

Cited By ~ 6

Author(s):

John T Szilagyi ◽

Anastasia N Freedman ◽

Stewart L Kepper ◽

Arjun M Keshava ◽

Jackie T Bangma ◽

...

Keyword(s):

Chemokine Receptors ◽

Cell Types ◽

The United States ◽

Trophoblast Invasion ◽

Migration And Invasion ◽

Placental Blood ◽

Polyfluoroalkyl Substances ◽

The Stability ◽

Trophoblast Migration

Abstract Per- and polyfluoroalkyl substances (PFAS) are used as industrial surfactants and chemical coatings for household goods such as Teflon. Despite regulatory efforts to phase out legacy PFAS, they remain detectable in drinking water throughout the United States. This is due to the stability of legacy PFAS and the continued use of replacement compounds. In humans, PFAS have been detected in placenta and cord blood and are associated with low birth weight and preeclampsia risk. Preeclampsia is a leading cause of maternal mortality and is driven by insufficient endometrial trophoblast invasion, resulting in poor placental blood flow. PFAS alter invasion of other cell types, but their impact on trophoblasts is not understood. We therefore assessed the effects of PFAS on trophoblast migration, invasion, and gene expression in vitro. Trophoblast migration and invasion were assessed using a modified scratch assay in the absence or presence of Matrigel, respectively. Treatment with perfluorooctanoic sulfate (PFOS), perfluorooctanoic acid (PFOA), and GenX (1000 ng/ml) each decreased trophoblast migration over 24 h. However, only GenX (1000 ng/ml) significantly inhibited trophoblast invasion. Treatment with PFOS, PFOA, and GenX also decreased trophoblast expression of chemokines (eg, CCL2), chemokine receptors (eg, CCR4), and inflammatory enzymes (eg, ALOX15) involved in migration. Inhibition of chemokine receptors with pertussis toxin (10 ng/ml), a G-protein inhibitor, inhibited trophoblast migration similar to the PFAS. Taken together, PFAS decrease trophoblast migration, invasion, and inflammatory signaling. By understanding the mechanisms involved, it may be possible to identify the biological and exposure factors that contribute to preeclampsia.

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275. Neuronal regeneration peptide (NRP): a novel trophoblast migration and survival enhancing factor

Reproduction Fertility and Development ◽

10.1071/srb05abs275 ◽

2005 ◽

Vol 17 (9) ◽

pp. 113

Author(s):

A. Singh ◽

J. Keelan ◽

F. Sieg

Keyword(s):

Biological Activities ◽

Trophoblast Invasion ◽

Migration And Invasion ◽

Mitochondrial Activity ◽

Placental Development ◽

Paracrine Factors ◽

Tnf Α ◽

Migration Assays ◽

Choriocarcinoma Cells ◽

Trophoblast Migration

Autocrine and paracrine factors regulate survival, proliferation, migration and invasion of placental cytotrophoblasts cells. While trophoblast migration appears to be tightly controlled, the nature of the chemoattractive factors that facilitate and direct trophoblast invasion remains undefined. Our group recently discovered a chemottractive factor (NRP) that exerts its biological activities on the CNS. Studies of NRP actions reveal extensive influences on postnatal neuronal migration, differentiation and survival. Based on the neuronal activities of NRPs and parallels between neuronal and placental cell behavior patterns, we speculated that NRPs could be involved in placental development. Migration assays were performed over 22 h using Boyden chambers pre-coated with NRP and laminin (n = 6 chambers per condition), to test the effect of NRPs on trophoblast migration. CTBs were isolated from term placentas by trypsin digestion and Percoll purification, and experiments were conducted within 6 h. Trophoblasts were seeded into the inner chamber (50000 cells/well) in M199 media supplemented with 10% FBS and antibiotics. Total number of cells migrating was counted. Migration was increased by 95 ± 14 % (mean ± SEM) in the presence of 100 fM NRP (P = 0.0016, t-test) compared to controls (bovine serum albumin). Survival assays were also performed using both primary trophoblasts and Jar choriocarcinoma cells. Apoptosis over 48 h, induced by treatment with TNF-α (5 ng/mL) and IFN-γ (100 U/mL), was completely abrogated by 10 pM NRP in Jar cells and 1pM in primary trophoblasts, as judged by MTT assay (mitochondrial activity). Inadequate placentation is implicated in the pathogenesis of a number of serious pregnancy disorders such as preeclampsia. Our findings suggest important roles for NRPs in regulating trophoblast migration and survival. The possibility that defective NRP actions may be involved in various placental pathologies, or that NRPs could be used pharmacologically to augment placentation, remain to be explored.

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Potential Role of microRNA-183 as a Tumor Suppressor in Hepatocellular Carcinoma

Cellular Physiology and Biochemistry ◽

10.1159/000495825 ◽

2018 ◽

Vol 51 (5) ◽

pp. 2065-2072 ◽

Cited By ~ 3

Author(s):

Wei Bian ◽

Hongfei Zhang ◽

Miao Tang ◽

Shaojun Zhang ◽

Lichao Wang ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Hepg2 Cells ◽

Molecular Mechanisms ◽

The Cancer Genome Atlas ◽

Migration And Invasion ◽

Metastatic Progression ◽

Mesenchymal Transition ◽

Reporter Assays

Background/Aims: Disseminated tumors, known as metastases, are responsible for ninety-percent of mortality due to cancer. Epithelial to mesenchymal transition, a phenomenon required for morphological conversion of non-motile discoid shaped epithelial cells to highly motile spindle-shaped mesenchymal cells, is thought to be a pre-requisite for metastatic progression. Metastasis-associated 1 (MTA1) protein is a prime inducer of EMT and metastatic progression in all solid tumors including hepatocellular carcinoma (HCC). However, the molecular mechanisms that regulate the expression and function of MTA1 in HCC have not been elucidated. Methods: In silico prediction algorithms were used to find microRNAs (miRNAs) that may target MTA1. We examined the relationship between the expression of MTA1 and miR-183 using quantitative real time PCR. We also determined the levels of the MTA1 protein using immunohistochemistry. Reporter assays, in the presence and absence of the miR-183 mimic, were used to confirm MTA1 as a bona fide target of miR183. The effect of miR-183 on HCC pathogenesis was determined using a combination of in vitro migration and invasion assay, together with in vivo xenograft experiments. The correlation between miR-183 and MTA1 expression was also studied in samples from HCC patients, and in The Cancer Genome Atlas dataset. Results: Analysis of the sequence database revealed that MTA1 is a putative target of miR-183. MTA1 protein and RNA expression showed opposite trends to miR-183 expression in breast, renal, prostate, and testicular tissue samples from cancer patients, and in the metastatic HCC cell line HepG2. An inverse correlation was also observed between MTA1 (high) and miR-183 (low) expression within samples from HHC patients and in the TCGA dataset. Reporter assays in HepG2 cells showed that miR-183 could inhibit translation of a reporter harboring the wild-type, but not the mutant miR-183 3’-untranslated region (UTR). In addition, miR-183 significantly inhibited in vitro migration and invasion in HepG2 cells, and in vivo hepatic metastasis. Conclusion: Our results reveal a novel post-transcriptional regulatory mechanism for MTA1 expression via miR-183, which is suppressed during HCC pathogenesis.

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BAG3 contributes to HGF-mediated cell proliferation, migration, and invasion via the Egr1 pathway in gastric cancer

Tumori Journal ◽

10.1177/0300891618811274 ◽

2018 ◽

Vol 105 (1) ◽

pp. 63-75

Author(s):

Jae Chang Lee ◽

Sung Ae Koh ◽

Kyung Hee Lee ◽

Jae-Ryong Kim

Keyword(s):

Gastric Cancer ◽

Cell Proliferation ◽

Cell Invasion ◽

Molecular Mechanisms ◽

Migration And Invasion ◽

Human Gastric Cancer ◽

Protein Levels ◽

Dependent Pathway

Introduction: Bcl2-associated athanogene 3 (BAG3) is elevated in several types of cancers. However, the role of BAG3 in progression of gastric cancer is unknown. Therefore, the present study aims to find out the role of BAG3 in hepatocyte growth factor (HGF)–mediated tumor progression and the molecular mechanisms by which HGF regulates BAG3 expression. Methods: BAG3 mRNA and protein were measured using reverse transcription polymerase chain reaction and Western blot in the 2 human gastric cancer cell lines, NUGC3 and MKN28, treated with or without HGF. The effects of BAG3 knockdown on cell proliferation, cell invasion, and apoptosis were analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the in vitro 2-chamber invasion assay, and flow cytometry in BAG3 short hairpin RNA (shRNA)–transfected cells and control cells. The signaling pathways involved in BAG3 that are regulated by HGF were analyzed. The chromatin immunoprecipitation assay was used to determine binding of Egr1 to the BAG3 promoter. Results: BAG3 mRNA and protein levels were increased following treatment with HGF. HGF-mediated BAG3 upregulation increased cell proliferation and cell invasion; however, it decreased apoptosis. HGF-mediated BAG3 upregulation is regulated by an ERK and Egr1-dependent pathway. BAG3 may have an important role in HGF-mediated cell proliferation and metastasis in gastric cancer through an ERK and Egr1-dependent pathway. Conclusion: This pathway may provide novel therapeutic targets and provide information for further identification of other targets of therapeutic significance in gastric cancer.

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HO-1/CO Maintains Intestinal Barrier Integrity through NF-κB/MLCK Pathway in Intestinal HO-1-/- Mice

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/6620873 ◽

2021 ◽

Vol 2021 ◽

pp. 1-19

Author(s):

Zhenling Zhang ◽

Lijing Zhang ◽

Qiuping Zhang ◽

Bojia Liu ◽

Fang Li ◽

...

Keyword(s):

Molecular Mechanisms ◽

Intestinal Barrier ◽

Heme Oxygenase 1 ◽

Zinc Protoporphyrin ◽

Barrier Integrity ◽

Tnf Α ◽

Intestinal Barrier Integrity ◽

Deficient Mice

Background. Intestinal barrier injury is an important contributor to many diseases. We previously found that heme oxygenase-1 (HO-1) and carbon monoxide (CO) protect the intestinal barrier. This study is aimed at elucidating the molecular mechanisms of HO-1/CO in barrier loss. Materials and Methods. We induced gut leakiness by injecting carbon tetrachloride (CCl4) to wildtype or intestinal HO-1-deficient mice. In addition, we administrated tumor necrosis factor-α (TNF-α) to cells with gain- or loss-of-HO-1 function. The effects of HO-1/CO maintaining intestinal barrier integrity were investigated in vivo and in vitro. Results. Cobalt protoporphyrin and CO-releasing molecule-2 alleviated colonic mucosal injury and TNF-α levels; upregulated tight junction (TJ) expression; and inhibited epithelial IκB-α degradation and phosphorylation, NF-κB p65 phosphorylation, long MLCK expression, and MLC-2 phosphorylation after administration of CCl4. Zinc protoporphyrin completely reversed these effects. These findings were further confirmed in vitro, using Caco-2 cells with gain- or loss-of-HO-1-function after TNF-α. Pretreated with JSH-23 (NF-κB inhibitor) or ML-7 (long MLCK inhibitor), HO-1 overexpression prevented TNF-α-induced TJ disruption, while HO-1 shRNA promoted TJ damage even in the presence of JSH-23 or ML-7, thus suggesting that HO-1 dependently protected intestinal barrier via the NF-κB p65/MLCK/p-MLC-2 pathway. Intestinal HO-1-deficient mice further demonstrated the effects of HO-1 in maintaining intestinal barrier integrity and its relative mechanisms. Alleviated hepatic fibrogenesis and serum ALT levels finally confirmed the clinical significance of HO-1/CO repairing barrier loss in liver injury. Conclusion. HO-1/CO maintains intestinal barrier integrity through the NF-κB/MLCK pathway. Therefore, the intestinal HO-1/CO-NF-κB/MLCK system is a potential therapeutic target for diseases with a leaky gut.

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PLAU1 Facilitated Proliferation, Invasion, and Metastasis via Interaction With MMP1 in Head and Neck Squamous Carcinoma

Frontiers in Oncology ◽

10.3389/fonc.2021.574260 ◽

2021 ◽

Vol 11 ◽

Author(s):

Kun Wu ◽

Yuan-Yuan Mao ◽

Nan-Nan Han ◽

Hanjiang Wu ◽

Sheng Zhang

Keyword(s):

Head And Neck ◽

Colony Formation ◽

Molecular Mechanisms ◽

Malignant Neoplasm ◽

Migration And Invasion ◽

High Morbidity ◽

Invasion And Metastasis ◽

Matrix Metalloproteinase 1

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common malignant neoplasm; it is associated with high morbidity and mortality. Thus, understanding the molecular mechanisms underlying its initiation and progression is critical for establishing the most appropriate treatment strategies. We found that urokinase-type plasminogen activator (PLAU1) was upregulated and associated with poor prognosis in HNSCC. Silencing of PLAU1 inhibited the proliferation, colony-formation, migration, and invasion abilities of HNSCC cells in vitro and reduced the expression of matrix metalloproteinase 1 (MMP1), whereas PLAU1 overexpression significantly enhanced the growth, the colony-formation, migration, and invasion abilities, and the xenograft tumor growth of HNSCC cells in vivo and increased the expression of MMP1. The Co-IP assay verified that PLAU1 interacted with MMP1. A positive correlation between PLAU1 and MMP1 expression was observed in HNSCC samples. si-RNAs against MMP1 reversed the aggressive effects of PLAU1 overexpression in HNSCC. Taken together, our data revealed that PLAU1 facilitated HNSCC cell proliferation, invasion, and metastasis via interaction with MMP1.

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LINC00839 Knockdown Restrains The Metastatic Behaviors of Nasopharyngeal Carcinoma By Sponging miR-454-3p

10.21203/rs.3.rs-606939/v1 ◽

2021 ◽

Author(s):

Feng Ying Zhang ◽

Xia Li ◽

Ting Ting Huang ◽

Mei Ling Xiang ◽

Lin Lin Sun ◽

...

Keyword(s):

Nasopharyngeal Carcinoma ◽

Underlying Mechanism ◽

Luciferase Reporter ◽

Regulatory Function ◽

Migration And Invasion ◽

Luciferase Reporter Gene ◽

Non Coding Rna ◽

Invasive Capacity

Abstract Background Long intergenic non-coding RNA 00839 (LINC00839) has been verified as a cancer-promoting gene in malignancies. However, the significance of LINC00839 in nasopharyngeal carcinoma (NPC) has yet to be elaborated, as well as its underlying mechanism.Methods LINC00839 and miR-454-3p relative expression levels in NPC cells were examined by qRT-PCR. The growth of cells was examined by CCK-8 and colony formation assays. Cell migration and invasion were examined by wound healing and Transwell experiment, respectively. The binding sequence of LINC00839 and miR-454-3p was confirmed by the luciferase reporter gene experiment. The regulatory function of LINC00839 and miR-454-3p on c-Met was investigated by western blot.Results Here, we revealed that LINC00839 was elevated in NPC. Both LINC00839 knockdown and upregulation of miR-454-3p suppressed NPC cells proliferation, invasive capacity and EMT in vitro. Besides, LINC00839 was validated as a miR-454-3p “sponge”, and upregulation of LINC00839 could reverse miR-454-3p-mediated functions in NPC C666-1 and SUNE-1 cells. Furthermore, c-Met was determined to be targeted by miR-454-3p. Notably, c-Met was downregulated by LINC00839 knockdown through sponging miR-454-3p. In vivo, LINC00839 knockdown resulted in a slower tumor growth.Conclusions Altogether, knockdown of LINC00839 inhibits the aggressive properties of NPC cells via sponging miR-454-3p and regulating c-Met.

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TheBDKRB2 +9/-9 Polymorphisms Influence Pro-Inflammatory Cytokine Levels in Knee Osteoarthritis by Altering TLR-2 Expression: Clinical and in Vitro Studies

Cellular Physiology and Biochemistry ◽

10.1159/000443072 ◽

2016 ◽

Vol 38 (3) ◽

pp. 1245-1256 ◽

Cited By ~ 2

Author(s):

Shuo Chen ◽

Lei Zhang ◽

Ruonan Xu ◽

Yunfan Ti ◽

Yunlong Zhao ◽

...

Keyword(s):

Knee Osteoarthritis ◽

Inflammatory Cytokine ◽

Molecular Mechanisms ◽

Enzyme Linked Immunosorbent Assay ◽

Mrna Levels ◽

Knee Oa ◽

Tnf Α ◽

Cytokine Levels ◽

Underlying Mechanisms

Background/Aims: The bradykinin B2 receptor (BDKRB2) +9/-9 gene polymorphisms have been shown to be associated with the susceptibility and severity of osteoarthritis (OA); however, the underlying mechanisms are unclear. In this study, we investigated the correlation between the BDKRB2 +9/-9 polymorphisms and pro-inflammatory cytokine levels in OA and the molecular mechanisms involved. Methods: A total of 156 patients with primary knee OA and 121 healthy controls were enrolled. The BDKRB2 +9/-9 polymorphisms were genotyped. The tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8 levels were determined using Enzyme-linked immunosorbent assay (ELISA). The toll-like receptor (TLR)-2 and TLR-4 mRNA levels were determined by quantitative real-time PCR. The basal and bradykinin-stimulated pro-inflammatory cytokine secretion in human OA synoviocytes and the involvement of TLR-2 and mitogen-activated protein kinases (MAPKs) were investigated. Results: The presence of -9 bp genotype is associated with higher TNF-α, IL-6, and IL-8 levels and higher TLR-2 expression in OA patients. The basal and bradykinin-induced TLR-2 expressions in human OA synoviocytes were significantly reduced by specific inhibitors of p38, JNK1/2, and ERK1/2. Both the B2 receptor antagonist MEN16132 and TLR-2 silencing inhibited IL-6 and IL-8 secretion in human OA synoviocytes. Conclusion: The data suggested that the BDKRB2 +9/-9 polymorphisms influence pro-inflammatory cytokine levels in knee osteoarthritis by altering TLR-2 expression.

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Integrin switching regulates normal trophoblast invasion

Development ◽

10.1242/dev.120.12.3657 ◽

1994 ◽

Vol 120 (12) ◽

pp. 3657-3666 ◽

Cited By ~ 11

Author(s):

C.H. Damsky ◽

C. Librach ◽

K.H. Lim ◽

M.L. Fitzgerald ◽

M.T. McMaster ◽

...

Keyword(s):

First Trimester ◽

Uterine Wall ◽

Trophoblast Invasion ◽

Collagen Type Iv ◽

Fibronectin Receptor ◽

Type Iv ◽

Invasive Capacity ◽

Functional Consequences

Cells invade extracellular matrices in a regulated manner at specific times and places during normal development. A dramatic example is trophoblast invasion of the uterine wall. Previous studies have shown that differentiation of trophoblasts to an invasive phenotype is accompanied by temporally and spatially regulated switching of their integrin repertoire. In the first trimester human placenta, alpha 6 integrins are restricted to cytotrophoblast (CTB) stem cells and downregulated in invasive CTBs, whereas alpha 5 beta 1 and alpha 1 beta 1 integrins are upregulated in differentiating and invasive CTBs. The goal of the present study was to determine whether these changes have functional consequences for CTB invasiveness. Using an in vitro invasion model, we determined first that aggregates of invading first trimester CTBs in vitro undergo the same pattern of integrin switching as was observed in situ, thereby validating the utility of the model. We then showed that antibody perturbation of interactions involving laminin or collagen type IV and their integrin alpha 1/beta 1 receptor inhibited invasion by CTBs, whereas perturbing interactions between fibronectin and the alpha 5/beta 1 fibronectin receptor accelerated invasion. Finally, we report that later gestation CTBs, which display greatly decreased invasive capacity, are also unable to upregulate alpha 1 beta 1 complexes, providing further evidence that this integrin is critical for CTB invasion. This gestational regulation is transcriptional. These data indicate that integrin switching observed during differentiation in situ has significant functional consequences for CTB invasion. The data suggest further that differentiating CTBs upregulate counterbalancing invasion-accelerating and invasion-restraining adhesion mechanisms. We propose that this contributes to regulating the depth of CTB invasion during normal implantation.

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Pharmacological Inhibition of Caspase-8 Suppresses Inflammation-Induced Angiogenesis in the Cornea

Biomolecules ◽

10.3390/biom10020210 ◽

2020 ◽

Vol 10 (2) ◽

pp. 210

Author(s):

Yunzhe Tian ◽

He Li ◽

Xiuxing Liu ◽

Lihui Xie ◽

Zhaohao Huang ◽

...

Keyword(s):

Molecular Mechanisms ◽

Therapeutic Potential ◽

Corneal Neovascularization ◽

Caspase 8 ◽

Inhibitory Effects ◽

Toll Like Receptor 4 ◽

Pharmacological Inhibition ◽

Cytokines And Chemokines ◽

Tnf Α

Inflammation-induced angiogenesis is closely related to many diseases and has been regarded as a therapeutic target. Caspase-8 has attracted increasing attention for its immune properties and therapeutic potential in inflammatory disorders. The aim of our study is to investigate the clinical application of pharmacological inhibition of caspase-8 and the underlying molecular mechanisms in inflammation-induced angiogenesis in the cornea. A model of alkali burn (AB)-induced corneal neovascularization (CNV) in C57BL/6 wild-type (WT) mice and toll-like receptor 4 knockout (Tlr4-/-) mice was used. We found that AB increased caspase-8 activity and the pharmacological inhibition of caspase-8 exerted substantial inhibitory effects on CNV, with consistent decreases in caspase-8 activity, inflammatory cell infiltration, macrophage recruitment and activation, VEGF-A, TNF-α, IL-1β, MIP-1, and MCP-1 expression in the cornea. In vitro, caspase-8 mediated TLR4–dependent chemokines and VEGF-A production by macrophages. The TLR4 knockout significantly alleviated CNV, suppressed caspase-8 activity and downregulated expression of inflammatory cytokines and chemokines after AB. Taken together, these findings provide the first demonstration that the pharmacological inhibition of caspase-8 suppresses inflammation-induced angiogenesis and support the use of a pharmacological caspase-8 inhibitor as a novel clinical treatment for CNV and other angiogenic disorders.

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