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In Vitro Exposure of Human Luteinized Mural Granulosa Cells to Dibutyl Phthalate Affects Global Gene Expression
Michal Adir1, Mali Salmon-Divon2, Catherine M H Combelles3
1Infertility and IVF Unit, Department of Obstetrics and Gynecology, Chaim Sheba Medical Center, Tel Hashomer and Sackler School of Medicine, Tel-Aviv University, Israel.
Abstract:
Exposure to dibutyl phthalate (DBP) is ubiquitous among women of reproductive age. Previous studies in animal models and in human cells in vitro have shown that exposure to DBP disrupts ovarian function. Here, we examined the effect of DBP on global gene expression in mural granulosa cells (MGCs) in vitro. Primary cultures of MGC obtained from 48 patients undergoing IVF were treated with increasing concentrations of DBP (0, 0.01, 0.1, 1, 10, or 100 µg/ml) for 48 h. Microarray analysis was used to identify genes exhibiting expression changes following DBP exposure. When compared with untreated cells, exposure to 100 µg/ml DBP resulted in significant differences in expression of 346 annotated genes (> 2-fold; q value < .05). Of them, 151 were upregulated and 195 downregulated. The main functional annotations affected by DBP were associated with cell cycle, mitosis, Rho GTPases, PLK1, Aurora B signaling pathways, and E2F-mediated regulation of DNA replication. No significant differences in gene expression were observed for the lower concentrations of DBP (0.01, 0.1, 1, and 10 µg/ml) compared with controls for both the microarray analysis and genes validated by quantitative real-time (qRT)-PCR. This study provides important molecular inputs on the effect of short-term DBP exposure on human MGCs in vitro. Our results indicate that acute treatment with high concentrations of DBP alters gene expression pathways mainly associated with the cell cycle.
Insights
High concentrations of dibutyl phthalate (DBP) significantly alter gene expression in human mural granulosa cells (MGCs), impacting cell cycle and mitosis pathways. Lower DBP levels showed no significant effects on gene expression.
Area of Science:
- Reproductive biology
- Environmental toxicology
- Molecular genetics
Background:
- Dibutyl phthalate (DBP) is a common environmental chemical.
- Previous research indicates DBP disrupts ovarian function in animal models and human cells.
- The molecular effects of DBP on human mural granulosa cells (MGCs) require further investigation.
Purpose of the Study:
- To investigate the impact of DBP on global gene expression in human MGCs in vitro.
- To identify specific genes and pathways affected by DBP exposure.
Main Methods:
- Primary human MGCs from 48 IVF patients were cultured.
- Cells were exposed to varying concentrations of DBP (0-100 µg/ml) for 48 hours.
- Microarray analysis and quantitative real-time PCR (qRT-PCR) were used to assess gene expression changes.
Main Results:
- Exposure to 100 µg/ml DBP significantly altered the expression of 346 genes (151 upregulated, 195 downregulated).
- Affected pathways included cell cycle, mitosis, Rho GTPases, and E2F-mediated DNA replication.
- Lower DBP concentrations (0.01-10 µg/ml) did not result in significant gene expression changes.
Conclusions:
- Acute exposure to high concentrations of DBP affects global gene expression in human MGCs.
- DBP primarily impacts pathways regulating the cell cycle and mitosis.
- These findings provide molecular insights into the effects of DBP on ovarian cells.
