Small interfering RNA-mediated Nrf2 gene knockdown enhances hirsutanols A-induced cytotoxicity in cancer cells.
- Author:
Jianguo MA
1
;
Houjin LI
;
Rong DENG
;
Gongkan FENG
;
Xiaofeng ZHU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; drug effects; Cell Line, Tumor; Cell Proliferation; drug effects; Colonic Neoplasms; metabolism; pathology; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Liver Neoplasms; metabolism; pathology; NF-E2-Related Factor 2; genetics; RNA, Small Interfering; Reactive Oxygen Species; metabolism; Sesquiterpenes; pharmacology
- From: Journal of Southern Medical University 2012;32(8):1093-1097
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of Nrf2 gene knockdown on hirsutanols A-induced cytotoxicity in cancer cells.
METHODSThe changes in the cell viability following treatment with different concentrations of hirsutanols A was detected by MTT assay, and the generation of reactive oxygen species (ROS) was assayed using flow cytometry. AnnexinV-FITC apoptosis kit was used to detect the cell apoptosis. Nrf2 protein expression in HepG2 and SW480 cells transfected with the siRNA targeting Nrf2 was analyzed with Western blotting.
RESULTSAt the concentrations of 1.25, 2.5, 5, 10, 20 and 40 µmol/L, hirsutanols A obviously inhibited the cell proliferation of human liver cancer HepG2 and colon cancer SW480 cells in a concentration-dependent manner. The levels of hydrogen peroxide increased rapidly after hirsutanols A treatment in both HepG2 (30 µmol/L) and SW480 (15 µmol/L) cells. Hirsutanols A also induced apoptosis of the two cells. Pretreatment with 5 mmol/L NAC totally inhibited apoptosis and ROS accumulation in the two cells induced by hirsutanols A. Transfection of HepG2 and SW480 cells with the siRNA caused a significant reduction in Nrf2 protein expression, which resulted in an increased sensitivity of the cells to hirsutanols A.
CONCLUSIONHirsutanols A can induce apoptosis in HepG2 and SW480 cells by promoting ROS production and up-regulating Nrf2 expression. Nrf2 knockdown by siRNA can increase the sensitivity of the cancer cells to hirsutanols A in vitro.
