TPOL triggers apoptosis with mitochondrial injury through activating a ROS-dependent p53/p21/p27/Rb/Bax/Cyto C/caspase-mediated signaling
10.3969/j.issn.1000-4718.2024.08.016
- VernacularTitle:TPOL通过激活ROS依赖的p53/p21/p27/Rb/Bax/Cyto C/caspase信号引发伴随线粒体损伤的细胞凋亡
- Author:
Zongwei CHENG
1
;
Boning ZENG
;
Feiyue XING
Author Information
1. 暨南大学免疫生物学系,组织移植与免疫研究中心,广东 广州 510632
- Keywords:
ethyl(2,4,6-trimethylbenzoyl)phenylphosphinate;
reactive oxygen species;
mitochondria;
apoptosis
- From:
Chinese Journal of Pathophysiology
2024;40(8):1488-1496
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To explore the influence of ethyl(2,4,6-trimethylbenzoyl)phenylphosphinate(TPOL)on cell apoptosis and its potential mechanism.METHODS:HEK293T cells sensitive to TPOL were treated with different concentrations of TPOL with or without exposure to light radiation,before treatment with various inhibitors,N-acetyl-L-cysteine(NAC),pifithrin-α and Z-DVED-FMK.Cell viability was measured by CCK-8 assay.Annexin V/propidium io-dide staining was used to count the number of apoptotic cells.DCFH-DA staining was used to detect reactive oxygen spe-cies(ROS)levels,and JC-1 staining was used to assess mitochondrial membrane potential by flow cytometry.The expres-sion of apoptosis-related proteins and cell cycle-regulated molecules was measured by Western blot.RESULTS:TPOL enhanced the apoptosis of HEK293T cells in a dose-dependent manner(P<0.05),with a decrease in Bcl-2 and increases in Bax and cytochrome C(Cyto C),followed by up-regulation of activated caspase-9 and caspase-3,and the cleavage of PARP(P<0.05).The TPOL-enhanced cleavage of caspase-3 and PARP was rescued by Z-DVED-FMK(P<0.01).TPOL also led to a rapid increase in ROS,a reduction in mitochondrial membrane potential,and the release of Cyto C(P<0.01),all of which could be reversed by the ROS scavenger NAC.Moreover,the TPOL-caused alterations in p21,p27,Rb,and CDK2 were also recovered by the p53 inhibitor pifithrin-α(P<0.05).The TPOL-induced changes in Bax,Bcl-2,cleaved caspase-9,activated caspase-3,and cleaved PARP were subsequently rescued by pretreatment with pifithrin-α(P<0.05).CONCLUSION:TPOL can induce cellular apoptosis with ROS-mediated mitochondrial membrane damage through the activation of a ROS-dependent p53/p21/p27/Rb/Bax/Cyto C/caspase-mediated signal axis.
