Gossypol acetic acid induces apoptosis in RAW264.7 cells via a caspase-dependent mitochondrial signaling pathway.
10.4142/jvs.2013.14.3.281
- Author:
Sijun DENG
1
;
Hui YUAN
;
Jine YI
;
Yin LU
;
Qiang WEI
;
Chengzhi GUO
;
Jing WU
;
Liyun YUAN
;
Zuping HE
Author Information
1. College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China. dykeyan3618@163.com
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
apoptosis;
gossypol acetic acid;
mitochondrial signaling pathway
- MeSH:
Animals;
Antineoplastic Agents, Phytogenic/*pharmacology;
Apoptosis/*drug effects;
Cell Line;
Cell Proliferation/*drug effects;
Dose-Response Relationship, Drug;
Gossypol/*analogs & derivatives/pharmacology;
Membrane Potential, Mitochondrial/*drug effects;
Mice;
Mice, Inbred BALB C;
Reactive Oxygen Species/*metabolism;
Signal Transduction/*drug effects
- From:Journal of Veterinary Science
2013;14(3):281-289
- CountryRepublic of Korea
- Language:English
-
Abstract:
To investigate the effects of gossypol acetic acid (GA) on proliferation and apoptosis of the macrophage cell line RAW264.7 and further understand the possible underlying mechanism responsible for GA-induced cell apoptosis, RAW264.7 cells were treated with GA (25~35 micromol/L) for 24 h and the cytotoxicity was determined by MTT assay, while apoptotic cells were identified by TUNEL assay, acridine orange/ethidium bromide staining and flow cytometry. Moreover, mitochondrial membrane potential (DeltaPsi(m)) with Rhodamine 123 and reactive oxygen species (ROS) with DCFH-DA were analyzed by fluorescence spectrofluorometry. In addition, the expression of caspase-3 and caspase-9 was assessed by Western Blot assay. Finally, the GA-induced cell apoptosis was evaluated by flow cytometry in the present of caspase inhibitors Z-VAD-FMK and Ac-LEHD-FMK, respectively. GA significantly inhibited the proliferation of RAW264.7 cells in a dose-dependent manner, and caused obvious cell apoptosis and a loss of DeltaPsi(m) in RAW264.7 cells. Moreover, the ROS production in cells was elevated, and the levels of activated caspase-3 and caspase-9 were up-regulated in a dose-dependent manner. Notably, GA-induced cell apoptosis was markedly inhibited by caspase inhibitors. These results suggest that GA-induced RAW264.7 cell apoptosis may be mediated via a caspase-dependent mitochondrial signaling pathway.
