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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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Related in: MedlinePlus

Characterization of HSD10-transfected cell lines. A. Empty vector (EV) and HSD10 overexpression (HSD10 ov) whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). B. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in EV and HSD10 ov cells. Scale bar in B: 30 μm. C. Quantification of HSD10 immunofluorescence staining (depicted in B) displayed as fold increase (n = 5). D. Control shRNA and HSD10 shRNA whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). E. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in control shRNA and HSD10 shRNA cells. Scale bar in E: 30 μm. F. Quantification of HSD10 immunofluorescence staining (depicted in E) displayed as fold increase (n = 5). Data presented as mean ± SE. *P < 0.01, **P < 0.001, ***P < 0.0001 verses control group.
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Fig1: Characterization of HSD10-transfected cell lines. A. Empty vector (EV) and HSD10 overexpression (HSD10 ov) whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). B. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in EV and HSD10 ov cells. Scale bar in B: 30 μm. C. Quantification of HSD10 immunofluorescence staining (depicted in B) displayed as fold increase (n = 5). D. Control shRNA and HSD10 shRNA whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). E. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in control shRNA and HSD10 shRNA cells. Scale bar in E: 30 μm. F. Quantification of HSD10 immunofluorescence staining (depicted in E) displayed as fold increase (n = 5). Data presented as mean ± SE. *P < 0.01, **P < 0.001, ***P < 0.0001 verses control group.

Mentions: After generating PC-12 cancer cell lines expressing either an empty vector (EV) or an HSD10 overexpression (HSD10 ov) vector, we first examined HSD10 protein expression levels in the cells using immunoblot analysis. HSD10 protein expression was increased by 3-fold in HSD10 ov cells compared to EV cells (Figure 1A). The level of HSD10 expression was further verified by immunofluorescence staining (Figure 1B). The intensity of HSD10 staining was significantly enhanced in HSD10 ov cells in comparison with EV cells (Figure 1C). In addition to confirming increased levels of HSD10 in the PC-12 overexpression cell lines, the immunofluorescence assay confirmed the localization of HSD10 to mitochondria as is depicted in the merged picture of Figure 1B.Figure 1


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)

Characterization of HSD10-transfected cell lines. A. Empty vector (EV) and HSD10 overexpression (HSD10 ov) whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). B. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in EV and HSD10 ov cells. Scale bar in B: 30 μm. C. Quantification of HSD10 immunofluorescence staining (depicted in B) displayed as fold increase (n = 5). D. Control shRNA and HSD10 shRNA whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). E. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in control shRNA and HSD10 shRNA cells. Scale bar in E: 30 μm. F. Quantification of HSD10 immunofluorescence staining (depicted in E) displayed as fold increase (n = 5). Data presented as mean ± SE. *P < 0.01, **P < 0.001, ***P < 0.0001 verses control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4384325&req=5

Fig1: Characterization of HSD10-transfected cell lines. A. Empty vector (EV) and HSD10 overexpression (HSD10 ov) whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). B. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in EV and HSD10 ov cells. Scale bar in B: 30 μm. C. Quantification of HSD10 immunofluorescence staining (depicted in B) displayed as fold increase (n = 5). D. Control shRNA and HSD10 shRNA whole cell lysates were analyzed for HSD10 protein expression via immunoblotting. β-actin was used as the loading control, and HSD10 expression was normalized to actin (n = 4). E. Confocal microscopy demonstrating immunofluorescence staining of HSD10 alone (green), Mito Tracker Red alone (red), and these two antigens co-localized (yellow) in control shRNA and HSD10 shRNA cells. Scale bar in E: 30 μm. F. Quantification of HSD10 immunofluorescence staining (depicted in E) displayed as fold increase (n = 5). Data presented as mean ± SE. *P < 0.01, **P < 0.001, ***P < 0.0001 verses control group.
Mentions: After generating PC-12 cancer cell lines expressing either an empty vector (EV) or an HSD10 overexpression (HSD10 ov) vector, we first examined HSD10 protein expression levels in the cells using immunoblot analysis. HSD10 protein expression was increased by 3-fold in HSD10 ov cells compared to EV cells (Figure 1A). The level of HSD10 expression was further verified by immunofluorescence staining (Figure 1B). The intensity of HSD10 staining was significantly enhanced in HSD10 ov cells in comparison with EV cells (Figure 1C). In addition to confirming increased levels of HSD10 in the PC-12 overexpression cell lines, the immunofluorescence assay confirmed the localization of HSD10 to mitochondria as is depicted in the merged picture of Figure 1B.Figure 1

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