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Oxidative Stress Alters miRNA and Gene Expression Profiles in Villous First Trimester Trophoblasts.

Cross CE, Tolba MF, Rondelli CM, Xu M, Abdel-Rahman SZ - Biomed Res Int (2015)

Bottom Line: Cytotoxicity, determined using the SRB assay, was used to calculate the IC50 of H2O2.H2O2 exerted a concentration- and time-dependent cytotoxicity on 3A trophoblast cells.Tool for annotations of microRNAs (TAM) analysis indicated that these altered miRNAs fall into 43 clusters and 34 families, with 41 functions identified.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, The University of Texas Medical Branch, Galveston, TX 77555-1066, USA.

ABSTRACT
The relationship between oxidative stress and miRNA changes in placenta as a potential mechanism involved in preeclampsia (PE) is not fully elucidated. We investigated the impact of oxidative stress on miRNAs and mRNA expression profiles of genes associated with PE in villous 3A first trimester trophoblast cells exposed to H2O2 at 12 different concentrations (0-1 mM) for 0.5, 4, 24, and 48 h. Cytotoxicity, determined using the SRB assay, was used to calculate the IC50 of H2O2. RNA was extracted after 4 h exposure to H2O2 for miRNA and gene expression profiling. H2O2 exerted a concentration- and time-dependent cytotoxicity on 3A trophoblast cells. Short-term exposure of 3A cells to low concentration of H2O2 (5% of IC50) significantly altered miRNA profile as evidenced by significant changes in 195 out of 595 evaluable miRNAs. Tool for annotations of microRNAs (TAM) analysis indicated that these altered miRNAs fall into 43 clusters and 34 families, with 41 functions identified. Exposure to H2O2 altered mRNA expression of 22 out of 84 key genes involved in dysregulation of placental development. In conclusion, short-term exposure of villous first trimester trophoblasts to low concentrations of H2O2 significantly alters miRNA profile and expression of genes implicated in placental development.

No MeSH data available.


Related in: MedlinePlus

Heat map representation for the differential expression of genes associated with PE after 4 h exposure to H2O2 in villous 3A trophoblasts. Gene expression is represented in the heat map in the color scale of −3.073–3.073 in green-red color scheme (n = 2). Genes evaluated and their locations on the heat map are depicted in the associated table above the heat map.
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fig3: Heat map representation for the differential expression of genes associated with PE after 4 h exposure to H2O2 in villous 3A trophoblasts. Gene expression is represented in the heat map in the color scale of −3.073–3.073 in green-red color scheme (n = 2). Genes evaluated and their locations on the heat map are depicted in the associated table above the heat map.

Mentions: In order to investigate the effect of oxidative stress on mRNA expression profile, we determined mRNA expression levels of 84 genes potentially involved in preeclamptic pregnancies. Villous 3A cells were mock-exposed in complete medium or exposed to 25 μM H2O2 in complete medium for 4 hours and total RNA was isolated using the Qiagen miRNeasy Mini Kit. The Qiagen RT2 Preeclampsia Array was used to evaluate the expression of these 84 genes after H2O2 exposure. As shown in Figure 3, of these 84 genes, 22 were up- or downregulated by at least twofold after H2O2 exposure. Of these, only the HBEGF gene was found to be upregulated while the other 21 genes were downregulated (Table 3).


Oxidative Stress Alters miRNA and Gene Expression Profiles in Villous First Trimester Trophoblasts.

Cross CE, Tolba MF, Rondelli CM, Xu M, Abdel-Rahman SZ - Biomed Res Int (2015)

Heat map representation for the differential expression of genes associated with PE after 4 h exposure to H2O2 in villous 3A trophoblasts. Gene expression is represented in the heat map in the color scale of −3.073–3.073 in green-red color scheme (n = 2). Genes evaluated and their locations on the heat map are depicted in the associated table above the heat map.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4538339&req=5

fig3: Heat map representation for the differential expression of genes associated with PE after 4 h exposure to H2O2 in villous 3A trophoblasts. Gene expression is represented in the heat map in the color scale of −3.073–3.073 in green-red color scheme (n = 2). Genes evaluated and their locations on the heat map are depicted in the associated table above the heat map.
Mentions: In order to investigate the effect of oxidative stress on mRNA expression profile, we determined mRNA expression levels of 84 genes potentially involved in preeclamptic pregnancies. Villous 3A cells were mock-exposed in complete medium or exposed to 25 μM H2O2 in complete medium for 4 hours and total RNA was isolated using the Qiagen miRNeasy Mini Kit. The Qiagen RT2 Preeclampsia Array was used to evaluate the expression of these 84 genes after H2O2 exposure. As shown in Figure 3, of these 84 genes, 22 were up- or downregulated by at least twofold after H2O2 exposure. Of these, only the HBEGF gene was found to be upregulated while the other 21 genes were downregulated (Table 3).

Bottom Line: Cytotoxicity, determined using the SRB assay, was used to calculate the IC50 of H2O2.H2O2 exerted a concentration- and time-dependent cytotoxicity on 3A trophoblast cells.Tool for annotations of microRNAs (TAM) analysis indicated that these altered miRNAs fall into 43 clusters and 34 families, with 41 functions identified.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, The University of Texas Medical Branch, Galveston, TX 77555-1066, USA.

ABSTRACT
The relationship between oxidative stress and miRNA changes in placenta as a potential mechanism involved in preeclampsia (PE) is not fully elucidated. We investigated the impact of oxidative stress on miRNAs and mRNA expression profiles of genes associated with PE in villous 3A first trimester trophoblast cells exposed to H2O2 at 12 different concentrations (0-1 mM) for 0.5, 4, 24, and 48 h. Cytotoxicity, determined using the SRB assay, was used to calculate the IC50 of H2O2. RNA was extracted after 4 h exposure to H2O2 for miRNA and gene expression profiling. H2O2 exerted a concentration- and time-dependent cytotoxicity on 3A trophoblast cells. Short-term exposure of 3A cells to low concentration of H2O2 (5% of IC50) significantly altered miRNA profile as evidenced by significant changes in 195 out of 595 evaluable miRNAs. Tool for annotations of microRNAs (TAM) analysis indicated that these altered miRNAs fall into 43 clusters and 34 families, with 41 functions identified. Exposure to H2O2 altered mRNA expression of 22 out of 84 key genes involved in dysregulation of placental development. In conclusion, short-term exposure of villous first trimester trophoblasts to low concentrations of H2O2 significantly alters miRNA profile and expression of genes implicated in placental development.

No MeSH data available.


Related in: MedlinePlus