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Overexpression of 17β-hydroxysteroid dehydrogenase type 10 increases pheochromocytoma cell growth and resistance to cell death.

Carlson EA, Marquez RT, Du F, Wang Y, Xu L, Yan SS - BMC Cancer (2015)

Bottom Line: Across disease states, increased HSD10 levels can have a profound and varied impact, such as beneficial in Parkinson's disease and harmful in Alzheimer's disease.In this study, we examined the tumor-promoting effect of HSD10 in pheochromocytoma cells.Our findings demonstrate that overexpression of HSD10 accelerates pheochromocytoma cell growth, enhances cell respiration, and increases cellular resistance to cell death induction.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology & Toxicology, University of Kansas, Lawrence, KS, 66047, USA. e086c574@ku.edu.

ABSTRACT

Background: 17β-hydroxysteroid dehydrogenase type 10 (HSD10) has been shown to play a protective role in cells undergoing stress. Upregulation of HSD10 under nutrient-limiting conditions leads to recovery of a homeostatic state. Across disease states, increased HSD10 levels can have a profound and varied impact, such as beneficial in Parkinson's disease and harmful in Alzheimer's disease. Recently, HSD10 overexpression has been observed in some prostate and bone cancers, consistently correlating with poor patient prognosis. As the role of HSD10 in cancer remains underexplored, we propose that cancer cells utilize this enzyme to promote cancer cell survival under cell death conditions.

Methods: The proliferative effect of HSD10 was examined in transfected pheochromocytoma cells by growth curve analysis and a xenograft model. Fluctuations in mitochondrial bioenergetics were evaluated by electron transport chain complex enzyme activity assays and energy production. Additionally, the effect of HSD10 on pheochromocytoma resistance to cell death was investigated using TUNEL staining, MTT, and complex IV enzyme activity assays.

Results: In this study, we examined the tumor-promoting effect of HSD10 in pheochromocytoma cells. Overexpression of HSD10 increased pheochromocytoma cell growth in both in vitro cell culture and an in vivo xenograft mouse model. The increases in respiratory enzymes and energy generation observed in HSD10-overexpressing cells likely supported the accelerated growth rate observed. Furthermore, cells overexpressing HSD10 were more resistant to oxidative stress-induced perturbation.

Conclusions: Our findings demonstrate that overexpression of HSD10 accelerates pheochromocytoma cell growth, enhances cell respiration, and increases cellular resistance to cell death induction. This suggests that blockade of HSD10 may halt and/or prevent cancer growth, thus providing a promising novel target for cancer patients as a screening or therapeutic option.

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Effect of HSD10 overexpression on cellular resistance to death-inducing stimuli. A-B. Densitometry of MTT reduction in EV and HSD10 ov cells treated with A) 0, 0.1, 0.25, 0.5, 0.75, and 1 mM H2O2 (n = 3 for all groups), and B) 0, 0.1, and 0.25 mM TBH for 24 hours (n = 3 for all groups). Results demonstrated that HSD10 ov cells were more resistant to oxidative stress-induced cell death. C-D. ETC. complex IV enzyme activity was assessed in EV and HSD10 ov cells treated with 0.75 mM H2O2 for C) 24 hours (n = 3), and D) 0, 1, 6, and 16 hours (n = 6 for all time points). Results, displayed as nmol/mg protein/min/ml, showed that complex IV activity is enhanced in HSD10 ov cells under an oxidative stress condition. E. Confocal microscopy demonstrating TUNEL staining of cells undergoing apoptosis (green), nuclear staining with DAPI (blue), and these two antigens co-localized (merged) in EV and HSD10 ov cells, treated with 0 mM and 0.75 mM H2O2 for 24 hours. Scale bar in E: 30 μm. F. Quantification of TUNEL staining (depicted in E) displayed as the percentage of TUNEL positive cells (n = 4). Data presented as mean ± SE. *P < 0.05, **P < 0.01, ***P < 0.0001 versus EV control group (A-D) and versus EV and HSD10 ov non-treatment groups (F); #P < 0.01 versus EV treatment group (F).
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Fig5: Effect of HSD10 overexpression on cellular resistance to death-inducing stimuli. A-B. Densitometry of MTT reduction in EV and HSD10 ov cells treated with A) 0, 0.1, 0.25, 0.5, 0.75, and 1 mM H2O2 (n = 3 for all groups), and B) 0, 0.1, and 0.25 mM TBH for 24 hours (n = 3 for all groups). Results demonstrated that HSD10 ov cells were more resistant to oxidative stress-induced cell death. C-D. ETC. complex IV enzyme activity was assessed in EV and HSD10 ov cells treated with 0.75 mM H2O2 for C) 24 hours (n = 3), and D) 0, 1, 6, and 16 hours (n = 6 for all time points). Results, displayed as nmol/mg protein/min/ml, showed that complex IV activity is enhanced in HSD10 ov cells under an oxidative stress condition. E. Confocal microscopy demonstrating TUNEL staining of cells undergoing apoptosis (green), nuclear staining with DAPI (blue), and these two antigens co-localized (merged) in EV and HSD10 ov cells, treated with 0 mM and 0.75 mM H2O2 for 24 hours. Scale bar in E: 30 μm. F. Quantification of TUNEL staining (depicted in E) displayed as the percentage of TUNEL positive cells (n = 4). Data presented as mean ± SE. *P < 0.05, **P < 0.01, ***P < 0.0001 versus EV control group (A-D) and versus EV and HSD10 ov non-treatment groups (F); #P < 0.01 versus EV treatment group (F).

Mentions: To assess resistance to cell death, we treated the PC-12 HSD10 overexpression cells with various concentrations of H2O2 and tert-butyl hydroperoxide (TBH) for 24 hours to stimulate oxidative stress conditions as cancer cells are typically exposed to higher oxidative stress levels [31]. While cell viability steadily decreased for both cell groups as the chemical dosage increased, HSD10 ov cells demonstrated significantly higher reduction of MTT at 0.75 and 1 mM concentrations of H2O2 compared to EV cells (Figure 5A). Treatment of the cells with TBH showed similar results, with HSD10 ov cells reducing considerably more MTT compared to EV cells at the lowest dosage of TBH given (0.1 mM, Figure 5B). Thus, while these two oxidative stressors reduce cell viability in both EV and HSD10 ov cells, PC-12 cells with HSD10 overexpression exhibited more resistance to chemical-induced oxidative stress.Figure 5


Overexpression of 17β-hydroxysteroid dehydrogenase type 10 increases pheochromocytoma cell growth and resistance to cell death.

Carlson EA, Marquez RT, Du F, Wang Y, Xu L, Yan SS - BMC Cancer (2015)

Effect of HSD10 overexpression on cellular resistance to death-inducing stimuli. A-B. Densitometry of MTT reduction in EV and HSD10 ov cells treated with A) 0, 0.1, 0.25, 0.5, 0.75, and 1 mM H2O2 (n = 3 for all groups), and B) 0, 0.1, and 0.25 mM TBH for 24 hours (n = 3 for all groups). Results demonstrated that HSD10 ov cells were more resistant to oxidative stress-induced cell death. C-D. ETC. complex IV enzyme activity was assessed in EV and HSD10 ov cells treated with 0.75 mM H2O2 for C) 24 hours (n = 3), and D) 0, 1, 6, and 16 hours (n = 6 for all time points). Results, displayed as nmol/mg protein/min/ml, showed that complex IV activity is enhanced in HSD10 ov cells under an oxidative stress condition. E. Confocal microscopy demonstrating TUNEL staining of cells undergoing apoptosis (green), nuclear staining with DAPI (blue), and these two antigens co-localized (merged) in EV and HSD10 ov cells, treated with 0 mM and 0.75 mM H2O2 for 24 hours. Scale bar in E: 30 μm. F. Quantification of TUNEL staining (depicted in E) displayed as the percentage of TUNEL positive cells (n = 4). Data presented as mean ± SE. *P < 0.05, **P < 0.01, ***P < 0.0001 versus EV control group (A-D) and versus EV and HSD10 ov non-treatment groups (F); #P < 0.01 versus EV treatment group (F).
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Related In: Results  -  Collection

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Fig5: Effect of HSD10 overexpression on cellular resistance to death-inducing stimuli. A-B. Densitometry of MTT reduction in EV and HSD10 ov cells treated with A) 0, 0.1, 0.25, 0.5, 0.75, and 1 mM H2O2 (n = 3 for all groups), and B) 0, 0.1, and 0.25 mM TBH for 24 hours (n = 3 for all groups). Results demonstrated that HSD10 ov cells were more resistant to oxidative stress-induced cell death. C-D. ETC. complex IV enzyme activity was assessed in EV and HSD10 ov cells treated with 0.75 mM H2O2 for C) 24 hours (n = 3), and D) 0, 1, 6, and 16 hours (n = 6 for all time points). Results, displayed as nmol/mg protein/min/ml, showed that complex IV activity is enhanced in HSD10 ov cells under an oxidative stress condition. E. Confocal microscopy demonstrating TUNEL staining of cells undergoing apoptosis (green), nuclear staining with DAPI (blue), and these two antigens co-localized (merged) in EV and HSD10 ov cells, treated with 0 mM and 0.75 mM H2O2 for 24 hours. Scale bar in E: 30 μm. F. Quantification of TUNEL staining (depicted in E) displayed as the percentage of TUNEL positive cells (n = 4). Data presented as mean ± SE. *P < 0.05, **P < 0.01, ***P < 0.0001 versus EV control group (A-D) and versus EV and HSD10 ov non-treatment groups (F); #P < 0.01 versus EV treatment group (F).
Mentions: To assess resistance to cell death, we treated the PC-12 HSD10 overexpression cells with various concentrations of H2O2 and tert-butyl hydroperoxide (TBH) for 24 hours to stimulate oxidative stress conditions as cancer cells are typically exposed to higher oxidative stress levels [31]. While cell viability steadily decreased for both cell groups as the chemical dosage increased, HSD10 ov cells demonstrated significantly higher reduction of MTT at 0.75 and 1 mM concentrations of H2O2 compared to EV cells (Figure 5A). Treatment of the cells with TBH showed similar results, with HSD10 ov cells reducing considerably more MTT compared to EV cells at the lowest dosage of TBH given (0.1 mM, Figure 5B). Thus, while these two oxidative stressors reduce cell viability in both EV and HSD10 ov cells, PC-12 cells with HSD10 overexpression exhibited more resistance to chemical-induced oxidative stress.Figure 5

Bottom Line: Across disease states, increased HSD10 levels can have a profound and varied impact, such as beneficial in Parkinson's disease and harmful in Alzheimer's disease.In this study, we examined the tumor-promoting effect of HSD10 in pheochromocytoma cells.Our findings demonstrate that overexpression of HSD10 accelerates pheochromocytoma cell growth, enhances cell respiration, and increases cellular resistance to cell death induction.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology & Toxicology, University of Kansas, Lawrence, KS, 66047, USA. e086c574@ku.edu.

ABSTRACT

Background: 17β-hydroxysteroid dehydrogenase type 10 (HSD10) has been shown to play a protective role in cells undergoing stress. Upregulation of HSD10 under nutrient-limiting conditions leads to recovery of a homeostatic state. Across disease states, increased HSD10 levels can have a profound and varied impact, such as beneficial in Parkinson's disease and harmful in Alzheimer's disease. Recently, HSD10 overexpression has been observed in some prostate and bone cancers, consistently correlating with poor patient prognosis. As the role of HSD10 in cancer remains underexplored, we propose that cancer cells utilize this enzyme to promote cancer cell survival under cell death conditions.

Methods: The proliferative effect of HSD10 was examined in transfected pheochromocytoma cells by growth curve analysis and a xenograft model. Fluctuations in mitochondrial bioenergetics were evaluated by electron transport chain complex enzyme activity assays and energy production. Additionally, the effect of HSD10 on pheochromocytoma resistance to cell death was investigated using TUNEL staining, MTT, and complex IV enzyme activity assays.

Results: In this study, we examined the tumor-promoting effect of HSD10 in pheochromocytoma cells. Overexpression of HSD10 increased pheochromocytoma cell growth in both in vitro cell culture and an in vivo xenograft mouse model. The increases in respiratory enzymes and energy generation observed in HSD10-overexpressing cells likely supported the accelerated growth rate observed. Furthermore, cells overexpressing HSD10 were more resistant to oxidative stress-induced perturbation.

Conclusions: Our findings demonstrate that overexpression of HSD10 accelerates pheochromocytoma cell growth, enhances cell respiration, and increases cellular resistance to cell death induction. This suggests that blockade of HSD10 may halt and/or prevent cancer growth, thus providing a promising novel target for cancer patients as a screening or therapeutic option.

Show MeSH
Related in: MedlinePlus