Integration of transcriptome and proteome profiles in placenta accreta reveals trophoblast over-migration as the underlying pathogenesis

Abstract Background Placenta accreta (PA) is a major cause of maternal morbidity and mortality in modern obstetrics, few studies have explored the underlying molecular mechanisms. Methods In our study, transcriptome and proteome profiling were performed in placental tissues from ten participants including five cases each in the PA and control groups to clarify the pathogenesis of PA. Results We identified differential expression of 37,743 transcripts and 160 proteins between the PA and control groups with an overlap rate of 0.09%. The 33 most-significant transcripts and proteins were found and further screened and analyzed. Adhesion-related signature, chemotaxis related signatures and immune related signature were found in the PA group and played a certain role. Sum up two points, three significant indicators, methyl-CpG-binding domain protein 2 (MeCP2), podocin (PODN), and apolipoprotein D (ApoD), which participate in “negative regulation of cell migration”, were downregulated at the mRNA and protein levels in PA group. Furthermore, transwell migration and invasion assay of HTR-8/SVneo cell indicated the all of them impaired the migration and invasion of trophoblast. Conclusion A poor correlation was observed between the transcriptome and proteome data and MeCP2, PODN, and ApoD decreased in transcriptome and proteome profiling, resulting in increased migration of trophoblasts in the PA group, which clarify the mechanism of PA and might be the biomarkers or therapy targets in the future.

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Conjugated Linoleic Acid Causes a Marked Increase in Liver α-Tocopherol and Liver α-Tocopherol Transfer Protein in C57BL/6 J Mice

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831/a000007 ◽

2010 ◽

Vol 80 (1) ◽

pp. 65-73 ◽

Cited By ~ 10

Author(s):

Pei-Min Chao ◽

Wan-Hsuan Chen ◽

Chun-Huei Liao ◽

Huey-Mei Shaw

Keyword(s):

Antioxidant Enzymes ◽

Linoleic Acid ◽

Conjugated Linoleic Acid ◽

Thiobarbituric Acid ◽

Control Group ◽

Thiobarbituric Acid Reactive Substances ◽

Control Groups ◽

Protein Levels ◽

Transfer Protein ◽

And Control

Conjugated linoleic acid (CLA) is a collective term for the positional and geometric isomers of a conjugated diene of linoleic acid (C18:2, n-6). The aims of the present study were to evaluate whether levels of hepatic α-tocopherol, α-tocopherol transfer protein (α-TTP), and antioxidant enzymes in mice were affected by a CLA-supplemented diet. C57BL/6 J mice were divided into the CLA and control groups, which were fed, respectively, a 5 % fat diet with or without 1 g/100 g of CLA (1:1 mixture of cis-9, trans-11 and trans-10, cis-12) for four weeks. α-Tocopherol levels in plasma and liver were significantly higher in the CLA group than in the control group. Liver α-TTP levels were also significantly increased in the CLA group, the α-TTP/β-actin ratio being 2.5-fold higher than that in control mice (p<0.01). Thiobarbituric acid-reactive substances were significantly decreased in the CLA group (p<0.01). There were no significant differences between the two groups in levels of three antioxidant enzymes (superoxide dismutase, glutathione peroxidase, and catalase). The accumulation of liver α-tocopherol seen with the CLA diet can be attributed to the antioxidant potential of CLA and the ability of α-TTP induction. The lack of changes in antioxidant enzyme protein levels and the reduced lipid peroxidation in the liver of CLA mice are due to α-tocopherol accumulation.

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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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Occult Placenta Accreta: The Missing Link in the Diagnosis of Abnormal Placentation

Pediatric and Developmental Pathology ◽

10.2350/06-10-0174.1 ◽

2007 ◽

Vol 10 (4) ◽

pp. 266-273 ◽

Cited By ~ 44

Author(s):

Jerzy Stanek ◽

Zarius Drummond

Keyword(s):

Placenta Accreta ◽

Histological Finding ◽

Cell Number ◽

Control Group ◽

Abnormal Placentation ◽

Control Groups ◽

Missing Link ◽

Intermediate Trophoblast ◽

And Control ◽

Extravillous Trophoblasts

Placenta creta (accreta, increta, or percreta) is a clinically symptomatic condition, usually diagnosed histologically on hysterectomy specimens. At a minimum, focal absence of decidua is the histological finding for this condition; however, excessive amounts of extravillous trophoblasts were recently documented on hysterectomy specimens. The histological finding of basal plate myometrial fibers (BPMF) without intervening decidua in spontaneously delivered placentas, which we term occult placenta accreta (OPA), is not infrequent, even in clinically asymptomatic cases. To prove that OPA is a missing link between normal placental implantation and clinical placenta accreta, CD146 immunohistochemical stains were performed on 25 sections of OPA (study group) and 25 placental sections without BPMF (control group). Implantation-site intermediate trophoblast (ISIT) cell number, thickness, and density were compared between the study and control groups. The ISIT micrometry thickness and cell number at BPMF sites were statistically significantly higher in OPA than in control group and same OPA placentas away from BPMF. There were no statistically significant differences in ISIT density. Therefore, although asymptomatic, OPA features the same histopathology as clinical placenta accreta and may share same pathogenesis, which may include decidual deficiency, abnormal trophoblast/ decidua interaction, and/or hypoxia.

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Overexpression of DDR1 Promotes Migration, Invasion, Though EMT-Related Molecule Expression and COL4A1/DDR1/MMP-2 Signaling Axis

Technology in Cancer Research & Treatment ◽

10.1177/1533033820973277 ◽

2020 ◽

Vol 19 ◽

pp. 153303382097327

Author(s):

Xin Xie ◽

Hongchao He ◽

Ning Zhang ◽

Xiaojing Wang ◽

Wenbin Rui ◽

...

Keyword(s):

Tyrosine Kinases ◽

Molecular Mechanisms ◽

Mrna Level ◽

Migration And Invasion ◽

Rt Pcr ◽

Protein Levels ◽

Signaling Axis ◽

Slug Expression ◽

Mrna And Protein Expression

Purpose: Discoidin domain receptor 1 (DDR1) belongs to a novel class of receptor tyrosine kinases. Previous evidence indicates that DDR1 overexpression promotes the aggressive growth of bladder cancer (BC) cells. This study aimed to investigate the molecular mechanisms by which DDR1 influences BC. Methods: DDR1 was transfected into human BC RT4 cells. DDR1, COL4A1, and MMP-2 expression in 30 BC tissues and paired adjacent tissues were examined by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry. Transwell assays were conducted to determine cell migration and invasion. RT-PCR and western blot (WB) were also used to measure the DDR1, COL4A1, MMP-2, and EMT-related gene (ZEB1 and SLUG) expression in RT4 cells after DDR1 overexpression. Results: COL4A1 and MMP-2 interacted with DDR1 in the PPI network. RT-PCR and immunohistochemistry results showed that both mRNA and protein levels of DDR1 and COL4A1 were significantly increased in BC tissue, while the expression of MMP-2 was increased only at the mRNA level ( P < 0.05). Overexpression of DDR1 in RT4 cells significantly promoted their migratory and invasive capabilities in vitro ( P < 0.05). Moreover, overexpression of DDR1 in RT4 cells increased the mRNA and protein expression of ZEB1, SLUG, COL4A1, and MMP-2 ( P < 0.01). DDR1-mediated migration and invasion of RT4 cells were reversed after COL4A1-siRNA treatment. Conclusion: DDR1 may be a potential therapeutic target in BC patients.

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Mesenchymal stem cell-derived exosomes protect trabecular meshwork from oxidative stress

Scientific Reports ◽

10.1038/s41598-021-94365-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ying-chao Li ◽

Juan Zheng ◽

Xi-zi Wang ◽

Xin Wang ◽

Wen-jing Liu ◽

...

Keyword(s):

Oxidative Stress ◽

Survival Rate ◽

Candidate Genes ◽

Inflammatory Cytokines ◽

Trabecular Meshwork ◽

Control Groups ◽

Beneficial Effects ◽

Protein Levels ◽

Trabecular Meshwork Cells ◽

And Control

AbstractThis study aims to investigate the beneficial effects of exosomes derived from bone marrow mesenchymal stem cells (BMSCs) on trabecular meshwork cells under oxidative stress and predict candidate genes associated with this process. Trabecular meshwork cells were pretreated with BMSC-derived exosomes for 24 h, and exposed to 0.1 mM H2O2 for 6 h. Survival rate of trabecular meshwork cells was measured with CCK-8 assay. Production of intracellular reactive oxygen species (iROS) was measured using a flow cytometer. RT-PCR and ELISA were used to detect mRNA and protein levels of inflammatory cytokines and matrix metalloproteinases (MMPs). Sequencing of RNA and miRNA for trabecular meshwork cells from Exo and control groups was performed on BGISEQ500 platform. Phenotypically, pretreatment of BMSC-derived exosomes improves survival rate of trabecular meshwork cells exposed to H2O2, reduces production of iROS, and inhibits expression of inflammatory cytokines, whereas increases expression of MMPs. There were 23 miRNAs, 307 lncRNAs, and 367 mRNAs differentially expressed between Exo and control groups. Exosomes derived from BMSCs may protect trabecular meshwork cells from oxidative stress. Candidate genes responsible for beneficial effects, such as DIO2 and HMOX1, were predicted.

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Targeting Hedgehog Pathway and DNA Methyltransferases in Uterine Leiomyosarcoma Cells

Cells ◽

10.3390/cells10010053 ◽

2020 ◽

Vol 10 (1) ◽

pp. 53

Author(s):

Natalia Garcia ◽

Ayman Al-Hendy ◽

Edmund C. Baracat ◽

Katia Candido Carvalho ◽

Qiwei Yang

Keyword(s):

Molecular Mechanisms ◽

Annexin V ◽

Dna Methyltransferases ◽

Migration And Invasion ◽

Uterine Leiomyosarcoma ◽

Uterine Smooth Muscle ◽

Protein Levels ◽

Gli Proteins ◽

Aggressive Tumor ◽

Nuclear Levels

Uterine leiomyosarcoma (LMS) is an aggressive tumor that presents a poor prognosis, high rates of recurrence, and metastasis. Because of its rarity, there is no information available concerning LMS molecular mechanisms of origin and development. Here, we assessed the expression profile of Hedgehog (HH) signaling pathway markers and the effects of their pharmacological inhibition on uterine smooth muscle (UTSM), leiomyoma, and LMS cells. Additionally, we also evaluated the effects of DNMTs inhibition on LMS cell behavior. Cell proliferation, migration and apoptosis rates were evaluated by MTT, Scratch, and Annexin V assays, respectively. RNA expression and protein levels were assessed by qRT-PCR and Western blot. We found that SMO and GLIs (1, 2, and 3) expression was upregulated in LMS cells, with increased nuclear levels of GLI proteins. Treatment with LDE225 (SMOi) and Gant61 (GLIi) resulted in a significant reduction in Glis protein levels in LMS (p < 0.05). Additionally, the expression of DNMT (1, 3a, and 3b), as well as GLI1 nuclear expression, was significantly decreased after treatment with HH inhibitor in LMS cells. Our results showed that blocking of SMO, GLI, and DNMTs is able to inhibit LMS proliferation, migration, and invasion. Importantly, the combination of those treatments exhibited a potentiated effect on LMS malignant features due to HH pathway deactivation.

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Nuclear Envelope Alterations in Myotonic Dystrophy Type 1 Patient-Derived Fibroblasts

International Journal of Molecular Sciences ◽

10.3390/ijms23010522 ◽

2022 ◽

Vol 23 (1) ◽

pp. 522

Author(s):

Diana Viegas ◽

Cátia D. Pereira ◽

Filipa Martins ◽

Tiago Mateus ◽

Odete A. B. da Cruz e Silva ◽

...

Keyword(s):

Nuclear Envelope ◽

Myotonic Dystrophy ◽

Molecular Mechanisms ◽

Myotonic Dystrophy Type 1 ◽

Myotonic Dystrophy Type ◽

Protein Levels ◽

Distal Muscle ◽

Multisystemic Disease ◽

And Control

Myotonic dystrophy type 1 (DM1) is a hereditary and multisystemic disease characterized by myotonia, progressive distal muscle weakness and atrophy. The molecular mechanisms underlying this disease are still poorly characterized, although there are some hypotheses that envisage to explain the multisystemic features observed in DM1. An emergent hypothesis is that nuclear envelope (NE) dysfunction may contribute to muscular dystrophies, particularly to DM1. Therefore, the main objective of the present study was to evaluate the nuclear profile of DM1 patient-derived and control fibroblasts and to determine the protein levels and subcellular distribution of relevant NE proteins in these cell lines. Our results demonstrated that DM1 patient-derived fibroblasts exhibited altered intracellular protein levels of lamin A/C, LAP1, SUN1, nesprin-1 and nesprin-2 when compared with the control fibroblasts. In addition, the results showed an altered location of these NE proteins accompanied by the presence of nuclear deformations (blebs, lobes and/or invaginations) and an increased number of nuclear inclusions. Regarding the nuclear profile, DM1 patient-derived fibroblasts had a larger nuclear area and a higher number of deformed nuclei and micronuclei than control-derived fibroblasts. These results reinforce the evidence that NE dysfunction is a highly relevant pathological characteristic observed in DM1.

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Mesenchymal Stem Cell-Derived Exosomes Protect Trabecular Meshwork from Oxidative Stress

10.21203/rs.3.rs-234332/v1 ◽

2021 ◽

Author(s):

Ying-chao Li ◽

Juan Zheng ◽

Xi-zi Wang ◽

Xin Wang ◽

Wen-jing Liu ◽

...

Keyword(s):

Oxidative Stress ◽

Survival Rate ◽

Candidate Genes ◽

Inflammatory Cytokines ◽

Trabecular Meshwork ◽

Control Groups ◽

Beneficial Effects ◽

Protein Levels ◽

Trabecular Meshwork Cells ◽

And Control

Abstract This study aims to investigate potential effects of exosomes derived from bone marrow mesenchymal stem cells (BMSCs) on trabecular meshwork cells under oxidative stress and predict candidate genes associated with this process. Trabecular meshwork cells were pretreated with BMSC-derived exosomes, and exposed to 0.1mM H2O2. Survival rate of trabecular meshwork cells was measured with CCK-8 assay. Production of iROS was measured using a flow cytometer. RT-PCR and ELISA were used to detect mRNA and protein levels of inflammatory cytokines and matrix metalloproteinases (MMPs). RNA sequencing for trabecular meshwork cells from Exo- and control groups was performed on BGISEQ500 + bgiseq-500 platform. Phenotypically, pretreatment of BMSC-derived exosomes improves survival rate of trabecular meshwork cells exposed to H2O2, reduces production of iROS, and inhibits expression of inflammatory cytokines, whereas increases expression of MMPs. There are 23 DEmiRNAs, 307 DElncRNAs, and 367 DEmRNAs differentially expressed between Exo- and control groups. Exosomes derived from BMSCs may protect trabecular meshwork cells from oxidative stress. Candidate genes responsible for beneficial effects ,such as DIO2 and HMOX1, such as DIO2 and HMOX1, are predicted.

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CDK1/FBXW7 Facilitates Degradation and Ubiquitination of MLST8 to Inhibit Progression of Renal Cell Carcinoma

10.21203/rs.3.rs-97872/v1 ◽

2020 ◽

Author(s):

Encheng Zhang ◽

Xiao Dong ◽

Jialiang Shao ◽

Xingyu Mu ◽

Siteng Chen ◽

...

Keyword(s):

Tumor Growth ◽

Renal Cancer ◽

Molecular Mechanisms ◽

Malignant Tumors ◽

Migration And Invasion ◽

Protein Levels ◽

Ubiquitin Proteasome ◽

Oncogenic Protein

Abstract Background: Recent studies have reported that MLST8 is upregulated in many malignant tumors. Nevertheless, the underlying molecular mechanisms is still unclear. The aim of this work was to investigate how MLST8 contribute to the development and progression of renal cancer (ccRCC).Methods: To identify molecular mediators of the oncogenic tumor function of MLST8, we analyzed a quantitative mass spectrometry by a previous study and checked the amino acid sequence in MLST8. Immunoprecipitation and Western Blotting were used to analyze the interaction between FBXW7 and MLST8. Transwell assays determined cell migration and invasion. In vivo experiments were performed to verify tumor growth. Immunohistochemistry was used to analyze protein levels in patients’ tumor samples. Results: MLST8 is an oncogenic protein in TCGA database and ccRCC clinical specimens. We also ascertain that MLST8 interacts with FBXW7, which was universally regarded as an E3 ubiquitin ligase. MLST8 can be degraded and ubiquitinated by tumor suppressor FBXW7. FBXW7 recognizes a consensus motif (T/S) PXX (S/T/D/E) of MLST8 and triggers MLST8 degradation via the ubiquitin-proteasome pathway. Strikingly, the activated CDK1 kinase engages in the MLST8 phosphorylation required for FBXW7-mediated degradation. In vitro and vivo assay, we further prove that MLST8 is an essential mediator of FBXW7 inactivation-induced tumor growth, migration, and invasion. Furthermore, MLST8 and FBXW7 protein are negatively correlated in human renal cancer specimens. Conclusions: Our findings suggest that MLST8 is a putative oncogene that functions via interaction with FBXW7, and inhibition MLST8 can be a potential future target in ccRCC treatment.

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Congenital cystic adenomatoid malformations of the lung: an epithelial transcriptomic approach

Respiratory Research ◽

10.1186/s12931-020-1306-5 ◽

2020 ◽

Vol 21 (1) ◽

Author(s):

Guillaume Lezmi ◽

Shamila Vibhushan ◽

Claudia Bevilaqua ◽

Nicolas Crapart ◽

Nicolas Cagnard ◽

...

Keyword(s):

Molecular Mechanisms ◽

Branching Morphogenesis ◽

Transcriptomic Analysis ◽

Differentially Expressed ◽

Protein Levels ◽

Redundant Genes ◽

And Control ◽

Laser Capture ◽

Congenital Cystic Adenomatoid Malformations

Abstract Background The pathophysiology of congenital cystic adenomatoid malformations (CCAM) of the lung remains poorly understood. Aim This study aimed to identify more precisely the molecular mechanisms limited to a compartment of lung tissue, through a transcriptomic analysis of the epithelium of macrocystic forms. Methods Tissue fragments displaying CCAM were obtained during planned surgical resections. Epithelial mRNA was obtained from cystic and normal areas after laser capture microdissection (LCM). Transcriptomic analyses were performed and the results were confirmed by RT-PCR and immunohistochemistry in independent samples. Results After controlling for RNA quality, we analysed the transcriptomes of six cystic areas and five control areas. In total, 393 transcripts were differentially expressed in the epithelium, between CCAM and control areas. The most highly redundant genes involved in biological functions and signalling pathways differentially expressed between CCAM and control epithelium included TGFB2, TGFBR1, and MAP 2 K1. These genes were considered particularly relevant as they have been implicated in branching morphogenesis. RT-qPCR analysis confirmed in independent samples that TGFBR1 was more strongly expressed in CCAM than in control tissues (p < 0.03). Immunohistochemistry analysis showed TGFBR1 (p = 0.0007) and TGFB2 (p < 0.02) levels to be significantly higher in the epithelium of CCAM than in that of control tissues. Conclusions This compartmentalised transcriptomic analysis of the epithelium of macrocystic lung malformations identified a dysregulation of TGFB signalling at the mRNA and protein levels, suggesting a possible role of this pathway in CCAM pathogenesis. Trial registration ClinicalTrials.gov Identifier: NCT01732185.

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