Celastrol induced hepatocellular carcinoma cell death via ROS-mediated DNA damage
- Author:
Jing MA
1
;
Hong-Lei WANG
1
;
Xin-Yu LI
1
;
Mei-Xiu LI
1
;
Yu-Jia LIU
1
;
Jing ZHOU
1
;
Jing MA
2
;
Zi-Xuan WANG
3
;
Yong WU
3
;
Guo-Dong LU
3
Author Information
- Publication Type:Journal Article
- Keywords: apoptosis; celastrol; DNA damage; hepatocellular carcinoma cells; oxidative damage; ROS
- From: Chinese Pharmacological Bulletin 2022;38(3):366-372
- CountryChina
- Language:Chinese
- Abstract: Aim To investigate the role of eelastrol in reactive oxygen species ( ROS) accumulation and DNA damage in hepatocellular carcinoma cells, and further investigate its effect on apoptosis induction in cancer cells.Methods Human liver cancer HepG2 and Huh7 cells were cultured with celastrol, then the morphological changes of cells were observed under microscope.MTT assay was employed to detect the proliferation of hepatocellular carcinoma cells, CM-H2DCFDA probe to detect intracellular ROS levels, and immunofluorescence to detect the expression level of -y-H2AX in celastrol-treated cells.Hoechst 33258 staining was used to observe nuclear condensation and fragmentation; Flow cytometry was used to evaluate cell death.J J Western blot was applied to measure the expression levels of -y-H2 AX, caspase-3, PARP and other proteins.Results Celastrol had a significant inhibitory effect on the proliferation of liver cancer cells in dose-dependent manner.Comparer] with the eontrol group, the eell viability was reduced and intracellular ROS level also increased significantly after celastrol treatment in a dose-dependent manner ( P < 0.05 ).With Hoechst staining, typical apoptotic characteristics such as nuclear chromatin condensation and fragmentation were observed in celastrol-treated cells.Western blot results showed that pro-form of caspase-3 significantly decreased, and the cleavage of PARP markedly increased by celastrol.After pretreatment with ROS inhibitor NAC, celastrol-mediated caspase-3 activation and PARP cleavage were significantly reversed ( P < 0.05 ).Conclusions Celastrol can induce apoptosis in hepatocellular carcinoma cells, and its anti-cancer effect is dependent on ROS-mediated DNA damage.
