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Effect of depletion of TET3 on gene expression of HTR8-SVneo trophoblast cells.

ABSTRACT: To investigate the function TET3 in the regulation of DNA methylation, we established HTR8-SVneo cell lines in which target gene has been knocked down by siRNA. We then performed gene expression profiling analysis using data obtained from RNA-seq for three replicates.

ORGANISM(S): Homo sapiens

PROVIDER: GSE211010 | GEO | 2022/08/14

REPOSITORIES: GEO

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Comparative proteome of the BeWo and HTR8/SVneo trophoblastic cell lines

Project description:Here, we have undertaken comparative label-free quantitative (LFQ) proteomic profiling of the proteins expressed in BeWo and HTR8/SVneo cell lines using a mass spectrometry approach. We have identified 1557 proteins in total. 915 proteins were expressed in both cell lines including various transcription factors, chaperones and transport proteins. A further 338 and 304 proteins were uniquely expressed in BeWo and HTR8/SVneo cells respectively. Our bioinformatics analysis reflects the known epithelial and mesenchymal phenotypes of these cell models with principal differences in GO observed in ‘Cell junction’, ‘Catenin complex’ as well as ‘Cell adhesion’ and ‘Cell differentiation’. Our novel comparative proteomic analysis of these trophoblastic cell lines will set the stage for the use of trophoblastic cell lines for the study of pregnancy disorders and further research focusing on placental function.

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Transcriptional change of HTR8/Svneo cells over expressed with PLAC8-EGFP

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Transcriptional change of HTR8/Svneo cells over expressed with PLAC8-EGFP

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Transcriptomic analysis of human extravillous trophoblast cells (HTR8/Svneo) cultured as 3D spheroids and 2D monolayers

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N6-methyladenosine (m6A) profiling and translatome of human trophoblast

Project description:To investigate the role of METTL3-mediated m6A modification and mRNA translation, we performed m6A-sequencing and Ribosome-nascent chain (RNC)-RNA sequencing in high glucose treated control or METTL3 knockdown HTR8/SVneo cells.

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Gene Expression Changes after Coumestrol Stimulation of Human Trophoblast Cells

Project description:Coumestrol is a potent isoflavone found in spinach and soy. To examine whether consuming large amounts of coumestrol during pregnancy affects the development of the placenta, human trophoblast cells (HTR8/Svneo) were treated with vehicle or coumestrol. Upon treatment, 3,079 genes were differentially regulated.

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PM2.5 leads to adverse pregnancy outcomes by inducing trophoblast oxidative stress and mitochondrial apoptosis via KLF9/CYP1A1 transcriptional axis

Project description:Epidemiological studies have demonstrated that fine particulate matter (PM2.5) is associated with adverse obstetric and postnatal metabolic health outcomes, but the mechanism remains unclear. This study aimed to investigate the toxicological pathways by which PM2.5 damaged placental trophoblasts in vivo and in vitro. We confirmed that PM2.5 induced adverse gestational outcomes such as increased fetal mortality rates, decreased fetal number and weight, damaged placental structure, and increased apoptosis of trophoblasts. Additionally, PM2.5 induced dysfunction of the trophoblast cell line HTR8/SVneo, including in its proliferation, apoptosis, invasion, migration and angiogenesis. Moreover, we comprehensively analyzed the transcriptional landscape of HTR8/SVneo cells exposed to PM2.5 through RNA-Seq and observed that PM2.5 triggered overexpression of pathways involved in oxidative stress and mitochondrial apoptosis to damage HTR8/SVneo cell biological functions through CYP1A1. Mechanistically, PM2.5 stimulated KLF9, a transcription factor identified as binding to CYP1A1 promoter region, which further modulated the CYP1A1-driven downstream phenotypes. Together, this study demonstrated that the KLF9/CYP1A1 axis played a crucial role in the toxic progression of PM2.5 induced adverse pregnancy outcomes, suggesting adverse effects of environmental pollution on pregnant females and putative targeted therapeutic strategies.

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5-Aza-dC treatment induces mesenchymal-to-epithelial transition in 1st trimester trophoblast cell line HTR8/SVneo.

Project description:Placental trophoblast invasion involves a cellular transition from epithelial to mesenchymal phenotype. Cytotrophoblasts undergo epithelial to mesenchymal transition (EMT) when differentiating into extravillous trophoblasts and gaining the capacity of invasion. In this research, we investigated the role of DNA methylation in trophoblasts during this EMT. First, using BeWo and HTR8/SVneo cell lines as models of cytotrophoblasts and extravillous trophoblasts, respectively, we analyzed the gene expression and DNA methylation status of the known epithelial marker genes, E-Cadherin and Cytokeratin7. We found that, in HTR8/SVneo cells, both genes were silenced and their promoters hypermethylated, as compared with the high-level gene expression and promoter hypomethylation observed in BeWo cells. This result suggests that dynamic DNA methylation of epithelial marker genes plays a critical role in the trophoblast EMT process. To verify these results, we treated HTR8/SVneo cells with 5-aza-dC, a known inhibitor of DNA methyltransferase, for three days. Five-Aza-dC treatment significantly increased the expression of epithelial marker genes and slightly decreased the expression of mesenchymal genes, as detected by qRT-PCR, immunocytochemistry and Western blot. Furthermore, 5-aza-dC treated HTR8/SVneo cells changed their morphology from mesenchymal into epithelial phenotype, indicating that 5-aza-dC induced mesenchymal to epithelial transition. Lastly, we examined the effect of 5-aza-dC on trophoblast migration and invasion capacity. We applied 5-aza-dC to HTR8/SVneo cells in trans-well cell migration and invasion assays and found that 5-aza-dC treatment decreased trophoblast migration and invasion capacity. In conclusion, DNA methylation of epithelial marker genes represents a molecular mechanism for the process of trophoblast EMT.

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Curcumin Protects Human Trophoblast HTR8/SVneo Cells from H2O2-Induced Oxidative Stress by Activating Nrf2 Signaling Pathway.

Project description:Pregnancy complications are associated with oxidative stress induced by accumulation of trophoblastic ROS in the placenta. We employed the human trophoblast HTR8/SVneo cell line to determine the effect of curcumin pre-treatment on H2O2-induced oxidative damage in HTR8/Sveo cells. Cells were pretreated with 2.5 or 5 ?M curcumin for 24 h, and then incubated with 400 ?M H2O2 for another 24 h. The results showed that H2O2 decreased the cell viability and induced excessive accumulation of reactive oxygen species (ROS) in HTR8/Sveo cells. Curcumin pre-treatment effectively protected HTR8/SVneo cells against oxidative stress-induced apoptosis via increasing Bcl-2/Bax ratio and decreasing the protein expression level of cleaved-caspase 3. Moreover, curcumin pre-treatment alleviated the excessive oxidative stress by enhancing the activity of antioxidative enzymes. The antioxidant effect of curcumin was achieved by activating Nrf2 and its downstream antioxidant proteins. In addition, knockdown of Nrf2 by Nrf2-siRNA transfection abolished the protective effects of curcumin on HTR8/SVneo cells against oxidative damage. Taken together, our results show that curcumin could protect HTR8/SVneo cells from H2O2-induced oxidative stress by activating Nrf2 signaling pathway.

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