Long-term in vitro exposure of human granulosa cells to the mixture of endocrine disrupting chemicals found in human follicular fluid disrupts steroidogenesis

Impact of In Vitro Long-Term Low-Level DEHP Exposure on Gene Expression Profile in Human Granulosa Cells

“…It has been reported that FSH and FOR (an adenylyl cyclase activator) could not induce aromatase (CYP19A1) mRNA expression or estradiol synthesis in HGrC1 cells. Progesterone production and the expression of STAR can be induced only after FOR stimulation [ 14 ]. Therefore, the effects of DEHP50 and DEHP250 in the current study were analyzed only at the level of basal estradiol production, whereas progesterone production was analyzed under the basal and FOR-stimulated conditions.…”

“…Human HGrC1 non-luteinized granulosa cells were kindly provided by Dr. A. Iwase (Nagoya University, Japan). The HGrC1 cells were cultured as described in [ 14 ]. For the long-term exposure study, three different cryopreserved stock vials of HGrC1 cells belonging to different passages (biological replicates) were thawed into three 25 cm 2 cell culture flasks and cultured for two weeks, after which the cells from each flask were divided into three 75 cm 2 flasks.…”

“…Western blot analysis was used to investigate protein expression after the long-term exposure of HGrC1 cells to DEHP. The analysis was performed as described in [ 14 ]. The dilution for the aromatase and STAR primary antibodies was 1:1000, and the dilution was 1:3000 for the GAPDH primary antibody.…”

“…To gain insight into the long-term effects of EDs in human cells, a limited number of studies employed immortalized cell lines that may be exposed to EDs for longer periods of time. Some of these studies revealed a distinct effect on steroidogenesis following a long-term exposure of human granulosa cells to the mixture of EDs [ 14 ], an ED-specific transcriptional reprograming in human breast cancer cells [ 15 ], dynamic, time-dependent changes in the response of human bronchial epithelial cells to the total particulate matter from a candidate modified-risk tobacco product [ 16 ], or a specific bisphenol-A-mediated effect in human vascular endothelial cells [ 17 ]. Moreover, long-term exposure studies also help in better understanding the biological effects of the relevant levels of several EDs in trophoblasts [ 18 ] or the molecular events involved in malignant transformation [ 19 ].…”

Paternal genetic effects of cadmium exposure during pregnancy on hormone synthesis disorders in ovarian granulosa cells of offspring

“…The mRNA expression levels of Star and Cyp17a1 decreased in the ovaries of neonatal mice exposed to phthalate diester [ 28 ]. The protein expression of Cyp19a1 was significantly reduced in human GCs HGrC1 after 2 weeks of exposure to a mixture composed of bisphenol A, PCBs, benzo [a] pyrene, and PFOS [ 29 ]. Previous studies have also observed significant decreases in Star and Cyp11a1 during hormone synthesis impairment in ovarian GCs of offspring induced by Cd exposure.…”