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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

Pie chart representation for miRNome array results after 4 h exposure to H2O2 in villous 3A trophoblasts.
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fig2: Pie chart representation for miRNome array results after 4 h exposure to H2O2 in villous 3A trophoblasts.

Mentions: To investigate the potential effect of oxidative stress on miRNA expression profile in villous 3A trophoblasts, cells were exposed to 25 μM H2O2 for 4 h in complete medium and total RNA (including miRNA) was isolated using the Qiagen miRNeasy Mini Kit. The Qiagen v16 miRNA Array was used to evaluate the expression of 1008 miRNAs after H2O2 exposure compared to unexposed control. In our study, 417 miRNAs were not expressed (unevaluable) in the tested cell line. Out of the 591 evaluable miRNAs, 195 were up- or downregulated by at least twofold after H2O2 exposure (Supplemental Table 1 available online at  http://dx.doi.org/10.1155/2015/257090). The majority of altered miRNAs (95.5%) were upregulated by at least 2-fold and only 4% were downregulated by at least 2-fold (Figure 2). Mir-21, -770, and -596 were downregulated by more than 5-fold, while mir-3907 was downregulated by more than 50-fold. Mir-637, mir-1911, mir-26b, mir-615, let-7a, and let-7f were upregulated more than 50-fold (Supplemental Table 1).


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)

Pie chart representation for miRNome array results after 4 h exposure to H2O2 in villous 3A trophoblasts.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig2: Pie chart representation for miRNome array results after 4 h exposure to H2O2 in villous 3A trophoblasts.
Mentions: To investigate the potential effect of oxidative stress on miRNA expression profile in villous 3A trophoblasts, cells were exposed to 25 μM H2O2 for 4 h in complete medium and total RNA (including miRNA) was isolated using the Qiagen miRNeasy Mini Kit. The Qiagen v16 miRNA Array was used to evaluate the expression of 1008 miRNAs after H2O2 exposure compared to unexposed control. In our study, 417 miRNAs were not expressed (unevaluable) in the tested cell line. Out of the 591 evaluable miRNAs, 195 were up- or downregulated by at least twofold after H2O2 exposure (Supplemental Table 1 available online at  http://dx.doi.org/10.1155/2015/257090). The majority of altered miRNAs (95.5%) were upregulated by at least 2-fold and only 4% were downregulated by at least 2-fold (Figure 2). Mir-21, -770, and -596 were downregulated by more than 5-fold, while mir-3907 was downregulated by more than 50-fold. Mir-637, mir-1911, mir-26b, mir-615, let-7a, and let-7f were upregulated more than 50-fold (Supplemental Table 1).

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