 |
 |
GEO help: Mouse over screen elements for information. |
|
| Status |
Public on Dec 31, 2015 |
| Title |
Oxidative stress alters miRNA and gene expression profiles in villous first trimester trophoblasts |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by RT-PCR
|
| Summary |
Oxidative stress plays a crucial role in preeclampsia (PE) pathogenesis. Evidence indicates altered microRNAs (miRNAs) expression in PE, however, the relationship between oxidative stress and miRNA changes in placenta as a potential mechanism involved in PE is not fully elucidated. In the present study, we investigate the impact of increased oxidative stress on miRNA and mRNA expression profiles of genes known to be associated with PE in villous 3A first trimester trophoblast cells. Cells were exposed to H2O2 at 10 different concentrations (0-1 mM) for 0.5, 4, 24, and 48 h. Cytotoxicity was determined using the SRB assay and data was used to calculate the IC50 of H2O2. Total RNA was extracted after short-term exposure (4 h) to H2O2 for miRNA 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 evaluatable miRNAs. Short-term exposure of villous first trimester trophoblasts to low concentrations of H2O2 significantly alters miRNA profile. Oxidative stress plays a crucial role in preeclampsia (PE) pathogenesis. Evidence indicates altered microRNAs (miRNAs) expression in PE, however, the relationship between oxidative stress and miRNA changes in placenta as a potential mechanism involved in PE is not fully elucidated. In the present study, we investigate the impact of increased oxidative stress on miRNA and mRNA expression profiles of genes known to be associated with PE in villous 3A first trimester trophoblast cells. Cells were exposed to H2O2 at 10 different concentrations (0-1 mM) for 0.5, 4, 24, and 48 h. Cytotoxicity was determined using the SRB assay and data was used to calculate the IC50 of H2O2. Total RNA was extracted after short-term exposure (4 h) to H2O2 for miRNA 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 evaluatable miRNAs. Short-term exposure of villous first trimester trophoblasts to low concentrations of H2O2 significantly alters miRNA profile.
|
| |
|
| Overall design |
Cells were grown to 90% confluency in 75 cm2 flasks and exposed to 25 µM H2O2 in complete medium for 4 h. Total RNA was isolated using the Qiagen miRNEasy Mini Kit and quality/quantity measured in the Molecular Genomics Core at UTMB. RNA was quantitated spectrophotometrically using a NanoDrop ND-1000 (NanoDrop Techniologies, DE). Quality of the purified RNA was assessed by visualization of 18S and 28S RNA bands using an Agilent BioAnalyzer 2100 (Agilent Technologies, CA). Resulting electroperograms were used in the calculation of the 28S/18S ratio and the RNA Integrity Number. Reverse transcription was carried out using either the miScript II RT kit or RT2 First Strand and subsequent SYBR green based real-time PCR on a BioRad Chromo4 Real-Time PCR Detector per manufacturer’s recommendation. The miRNA profile screening was performed using miScript Human miRNome PCR Array (MIHS-3216Z, Qiagen, Valencia, CA). Data was analyzed using the ΔΔCT method with the miScript miRNA PCR Array Data Analysis version 3.5 (SABiosciences, Valencia CA).
|
| |
|
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| |
| Submission date |
Dec 09, 2014 |
| Last update date |
Dec 31, 2015 |
| Contact name |
Mai Tolba |
| Organization name |
Faculty of Pharmacy, Ain Shams University
|
| Department |
Pharmacology and Toxicology
|
| Street address |
Abbasya
|
| City |
Cairo |
| State/province |
Non-US/Non-Canadian |
| ZIP/Postal code |
11566 |
| Country |
Egypt |
| |
|
| Platforms (1) |
| GPL19527 |
SA Biosciences Homo sapiens miRNome miRNA PCR Array v16 |
|
| Samples (2) |
|
| Relations |
| BioProject |
PRJNA269746 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE63999_fold-change.txt.gz |
5.0 Kb |
(ftp)(http) |
TXT |
| GSE63999_non-normalized.txt.gz |
5.5 Kb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
| Processed data are available on Series record |
|
|
|
|
 |
