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Updated: Mar 3, 2026

Malachite Green Assay for the Discovery of Heat-Shock Protein 90 Inhibitors
Published on: January 20, 2023
[Mechanism of heat shock protein 90 for regulating 26S proteasome in hyperthermia]
Qing-Rong Ma1, Pei-Zhi Yu, Fan Zhang
1Department of Thoracic Surgery, First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou 510080, China.
Objective:
To investigate the mechanism by which heat shock protein 90 (HSP90) regulates 26S proteasome in hyperthermia.
Methods:
Hyperthermic HepG2 cell models established by exposure of the cells to 42 degrees celsius; for 3, 6, 12, and 24 h were examined for production of reactive oxygen species (ROS) and cell proliferation, and the changes in Hsp90α and 26S proteasome were analyzed.
Results:
ROS production in the cells increased significantly after hyperthermia (F=28.958, P<0.001), and the cell proliferation was suppressed progressively as the heat exposure time extended (F=621.704, P<0.001). Hyperthermia up-regulated Hsp90α but decreased the expression level (F=164.174, P<0.001) and activity (F=133.043, P<0.001) of 26S proteasome. The cells transfected with a small interfering RNA targeting Hsp90α also showed significantly decreased expression of 26S proteasome (F=180.231, P<0.001).
Conclusion:
The intracellular ROS production increases as the hyperthermia time extends. Heat stress and ROS together cause protein denature, leading to increased HSP90 consumption and further to HSP90 deficiency for maintaining 26S proteasome assembly and stability. The accumulation of denatured protein causes unfolded protein reaction in the cells to eventually result in cell death.
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