Dehydroepiandrosterone inhibits cell proliferation and improves viability by regulating S phase and mitochondrial permeability in primary rat Leydig cells

Abstract

Dehydroepiandrosterone (DHEA) is widely used as a nutritional supplement and exhibits putative anti-aging properties. However, the molecular basis of the actions of DHEA, particularly on the biological characteristics of target cells, remain unclear. The aim of the current study was to investigate the effects of DHEA on cell viability, cell proliferation, cell cycle and mitochondrial function in primary rat Leydig cells. Adult Leydig cells were purified by Percoll gradient centrifugation, and cell proliferation was detected using a Click-iT® EdU Assay kit and cell cycle assessment performed using flow cytometry. Mitochondrial membrane potential was detected using JC-1 staining assay. The results of the current study demonstrate that DHEA decreased cell proliferation in a dose-dependent manner, whereas it improved cell viability in a time-dependent and dose-dependent manner. Flow cytometry analysis demonstrated that DHEA treatment increased the S phase cell population and decreased the G2/M cell population. Cyclin A and CDK2 mRNA levels were decreased in primary rat Leydig cells following DHEA treatment. DHEA treatment decreased the transmembrane electrical gradient in primary Leydig cells, whereas treatment significantly increased succinate dehydrogenase activity. These results indicated that DHEA inhibits primary rat Leydig cell proliferation by decreasing cyclin mRNA level, whereas it improves cells viability by modulating the permeability of the mitochondrial membrane and succinate dehydrogenase activity. These findings may demonstrate an important molecular mechanism by which DHEA activity is mediated.

Dehydroepiandrosterone Prevents H2O2-Induced BRL-3A Cell Oxidative Damage through Activation of PI3K/Akt Pathways rather than MAPK Pathways

“…Consistent with these changes in the active hormone content, a significant increase was observed in the protein expression of 3 β -HSD and 17 β -HSD after DHEA treatment. In agreement with our findings, DHEA has been shown to promote the testosterone and estradiol levels in both in vivo and in vitro settings [34, 35]. These findings can be explained through the process of bioconversion wherein 3 β -HSD catalyzes the rapid conversion of DHEA into androstenedione in peripheral organs such as the brain, liver, kidney, and gonads, after which androstenedione is converted into testosterone and estradiol by 17 β -HSD and aromatase, respectively [36].…”

Quinalizarin exerts an anti-tumour effect on lung cancer A549 cells by modulating the Akt, MAPK, STAT3 and p53 signalling pathways

“…In eukaryotes, mitosis is dependent on the completion of DNA synthesis (30). Checkpoints play important roles in the regulation of the cell cycle in eukaryotic cells, and abnormal regulation of these checkpoints frequently occurs in tumour cells (31). Cells may be arrested at G 0 /G 1 , S and G 2 /M phases of the cell cycle (32).…”

Clinical and Prognostic Significance of Cell Sensitivity to Chemotherapy Detected In Vitro on Treatment Response and Survival of Leukemia Patients

“…Sensitivity of tumor cells (IC 50 values) to a number of chemotherapeutic drugs was evaluated using the WST-test for all leukemia patients at diagnosis before the treatment start. The WST-assay method was applied in the study because, based on the estimation of mitochondrial dehydrogenase activity in cells, it is also useful as an indicator of cell viability and toxicity [32,33,34], and nowadays WST assay is widely used for the screening of drug responsiveness of cancer cells with different origins [35,36,37].…”