Evaluation of the effect of OPRM1 agonist on induction of cell death, apoptosis pattern, caspase 3 activity and the amount of reactive oxygen species in pituitary adenoma cells
نویسندگان: Fahimeh Taghavi1, Zahra Shahsavari2, Masoumeh Tavakoli-Yaraki1 ©
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Background and Aim
The pituitary gland is considered as one of the most important gland in the body which control body hemostasis, growth and proliferation. Cellular homeostasis is dependent on the balance between cell death and survival and this balance is controlled by intracellular complexing mechanisms. In this study, the effect of OPRM1 agonist on induction of cell death and activation of apoptosis, cell cycle distribution, the amount of reactive oxygen species in cells and the activity of caspases 3 activity in GH3 cells
The GH3 cells were treated with different concentrations of OPRM1 agonist at different time intervals and the rate of cell death was assessed by MTT assay. The Annexine V-FITC protocol by flow cytometry was used to measure the percentage of apoptotic, necrotic and viable cells. The activity of caspases 3 was examined by ELISA. The amount of intracellular ROS was assessed using fluorescence absorbance. The cell cycle distribution was assessed using PI protocol and flow cytometry.
Following activation of the OPRM1 receptor, the dose-dependent and time-dependent survival of GH3 cells showed a significant decrease, which was associated with an increase in the percentage of primary apoptosis in the cells. Also increased the percentage of cells in the Sub-G1 phase of the cell cycle, increased ROS production, and increased caspase-3 activity was observed in the GH3 cells. The results showed that simultaneous treatment with the receptor antagonist could restore the percentage of apoptosis, the percentage of cells in the Sub-G1 phase, the amount of ROS and caspase-3 activity induced by the agonist.
The OPRM1 agonist can induce apoptosis in GH3 cells in a concentration and time dependent manner by activating caspases and increasing the production of reactive oxygen species and showed the importance of the OPRM1 receptor in the pathogenesis of pituitary adenomas
Pituitary adenomas, OPRM1, apoptosis, cell cycle, caspase, intracellular free radicals
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