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Suppression of neutral cholesterol ester hydrolase activity by antisense DNA of hormone-sensitive lipase
J Osuga1, S Ishibashi, H Shimano
1Third Department of Internal Medicine, University of Tokyo, Japan.
Abstract:
In order to investigate the role of the hormone-sensitive lipase (HSL) gene in the activity of neutral cholesterol ester hydrolase (NCEH) from a molecular perspective, Chinese hamster ovary (CHO) cells were transfected with rat antisense hormone-sensitive lipase cDNA, and three different cell lines, designated as anti-HSL, were established. NCEH activity in anti-HSL cells was reduced to approximately 50% of that in control CHO cells. The concentration of cellular esterified cholesterol increased and the concentration of free cholesterol decreased in the anti-HSL cell lines. These results suggest that the HSL gene has a function of NCEH as well.
Insights
The hormone-sensitive lipase (HSL) gene plays a role in neutral cholesterol ester hydrolase (NCEH) activity. Reduced HSL gene expression in cells led to decreased NCEH activity and altered cholesterol levels.
Area of Science:
- Molecular biology
- Biochemistry
- Genetics
Background:
- Neutral cholesterol ester hydrolase (NCEH) is crucial for cellular cholesterol metabolism.
- The precise molecular mechanisms regulating NCEH activity are not fully understood.
- Hormone-sensitive lipase (HSL) is a known lipase, but its role in cholesterol ester hydrolysis requires further investigation.
Purpose of the Study:
- To investigate the molecular role of the hormone-sensitive lipase (HSL) gene in neutral cholesterol ester hydrolase (NCEH) activity.
- To determine if HSL gene expression influences NCEH function.
Main Methods:
- Chinese hamster ovary (CHO) cells were transfected with rat antisense hormone-sensitive lipase (HSL) cDNA.
- Three distinct anti-HSL cell lines were established.
- NCEH activity and cellular cholesterol concentrations (free and esterified) were measured in anti-HSL cells and compared to control CHO cells.
Main Results:
- NCEH activity was reduced by approximately 50% in anti-HSL cells compared to control cells.
- Cellular esterified cholesterol concentrations increased in anti-HSL cell lines.
- Cellular free cholesterol concentrations decreased in anti-HSL cell lines.
Conclusions:
- The HSL gene exhibits NCEH activity.
- HSL plays a significant role in regulating cellular cholesterol ester hydrolysis.
- These findings provide molecular insights into the function of HSL in cholesterol metabolism.