Effect Of Metformin, Simvastatin And Leptin On Proliferation And Secretion Of Adrenocortical Carcinoma H295R Cells

Part ?: Effect of Metformin on proliferation and secretion of adrenocortical carcinoma H295 R cellsObjective This study aims to investigate the effects of metformin on proliferation and secretion function of adrenocortical carcinoma H295 R cells.Methods H295 R cells were treated with different doses of metformin for 3 days or with 20 mmol/L metformine for different times.Cell proliferation was detected by MTS method.Cortisol and aldosterone in medium was determined by chemiluminescent method and radioimmunoassay,respectively.H295 R cells were treated with 20 mmol/L metformin for 24 hours,and m RNA expressions of 11 beta-hydroxylase(CYP11B1)and aldosterone synthase(CYP11B2)were detected by real-time quantitative PCR.?-catenin was observed by immunohistochemical staining.Results No differences of cell proliferation and secretion were detected between control group and metformin group with doses less than 5mmol/L.Metformin with large doses(? 10 mmol/L)significantly inhibited cell proliferation and secretion of cortisol and aldosterone in a dose-dependent pattern.Metformine(20 mmol/L)inhibited cell proliferation at 12 hours' treatment.And the inhibitory effects enhanced in a time-dependent manner.Compared with control group,metformin decreased secretion of cortisol and aldosterone,and m RNA expression of CYP11B1.?-catenin staining was not different between these two groups.Conclusions Metformin can inhibit cell proliferation and secretion of cortisol and aldosterone in H295R cells.Part ?: Effect of Simvastatin on proliferation and secretion of adrenocortical carcinoma H295 R cellsObjective To investigate the potential effects of simvastatin on proliferation and secretion of adrenocortical carcinoma H295 R cells.Methods The H295 R cells were divided into control group,Angiotensin II group,simvastatin group and Angiotensin II plus simvastatin group.Cell proliferation was detected by MTS method.Cortisol in medium was determined by chemiluminescent method,and aldosterone was determined by radioimmunoassay.The m RNA expressions of CYP11B1 and CYP11B2 were examined by real-time quantitative PCR.Results Angiotensin II had no effects on the cell proliferation,while simvastatin significantly inhibited cell proliferation.The inhibitory effect of simvastatin on proliferation was enhanced when simvastatin combined with angiotensin II(P < 0.05).Compared with control group,angiotensin II increased the secretion of cortisol and aldosterone,and the expression of CYP11B1 and CYP11B2.Simvastatin decreased cortisol secretion and CYP11B1 m RNA expression(P < 0.05).Simvastatin also inhibited angiotensin II-induced the secretion of cortisol and aldosterone,and the expression of CYP11B1 and CYP11B2 compared with Angiotensin II group(P < 0.05).Conclusions Simvastatin can inhibit angiotensin II-induced secretion of cortisol and aldosterone and the expression of CYP11B1 and CYP11B2 in H295 R cells.Simvastatin inhibits cell proliferation,which could be enhanced by simvastatin combined with angiotensin ?.Part ?: Effect of Leptin on proliferation and secretion of adrenocortical carcinoma H295 R cellsObjective To investigate the effects of leptin on proliferation and secretion of adrenocortical carcinoma H295 R cells.Methods The H295 R cells were divided into control group and leptin group,and treated for different times.Cell proliferation was detected by MTS method.H295 R cells were treated with 100 ng/m L leptin for 24 hours.Cortisol and aldosterone in medium was determined by chemiluminescent method and radioimmunoassay,respectively.Results Leptin did not affect cell proliferation from day 1 to day 7.Compared with control group,aldosterone increased in the leptin group(P < 0.05),while cortisol was unchanged.Conclusions Leptin significantly promotes aldosterone secretion,while has no effects on the cell proliferation and cortisol secretion in H295R cells.