Abstract
Thalictrum fortunei S. Moore, a perennial plant distributed in the southeastern part of China, has been used in Traditional Chinese Medicine for thousands of years for its antitumor, antibacterial and immunoregulatory effects. In order to investigate the active components and the mechanism of the anti-tumor effects of Thalictrum fortunei, the growth inhibitory effects of eight triterpenoids isolated from the aerial parts of the plant on tumor cell lines were examined by 3-(4,5)-dimethylthiazoy1-3,5-diphenyltetrazolium bromide (MTT) assay. The MTT-assay results showed that the inhibitory activity of 3-O-β-d-glucopyranosyl-(1→4)-β-d-fucopyranosyl(22S,24Z)-cycloart-24-en-3β,22,26-triol 26-O-β-d-glucopyranoside (1) was stronger than that of the other seven tested triterpenoids on human hepatoma Bel-7402 cell line (Bel-7402), human colon lovo cells (LoVo), human non-small cells lung cancer NCIH-460 cells (NCIH-460) and human gastric carcinoma SGC-7901 cells (SGC-7901) after 48 h treatment in vitro, with the IC50 values of 66.4, 84.8, 73.5, 89.6 μM, respectively. Moreover, the antitumor mechanism of compound 1 on Bel-7402 cell was explored through nucleus dyeing, fluorescence assay, flow cytometry and western blot. The flow cytometric analysis results revealed that compound 1 caused apoptosis and mitochondrial membrane potential (MMP) loss in Bel-7402 cells. A fluorescence assay indicated that intracellular reactive oxygen species (ROS) were markedly provoked by compound 1 treatment compared to control cells. Immunoblot results showed that compound 1 significantly increased the expression levels of cleaved caspase-3, P53 and Bax protein, and decreased the expression level of Bcl-2 protein. These findings indicate that compound 1 inhibits the growth activity of tumor cells, probably through the P53 protein-induced apoptosis pathway.
Highlights
Hepatocellular carcinoma is the eighth most common cancer over the World by number of cases and the third most common cause of death from cancer due to its very poor prognosis
The flow cytometric analysis results revealed that compound 1 caused apoptosis and mitochondrial membrane potential (MMP) loss in Bel-7402 cells
The aim of this study was to evaluate the cytotoxicity of eight triterpenoids from Thalictrum fortunei on SGC-7901 cells, Bel-7402 cell, lovo cells (LoVo) cells and NCIH-460 cells
Summary
Hepatocellular carcinoma is the eighth most common cancer over the World by number of cases and the third most common cause of death from cancer due to its very poor prognosis. Chemotherapy has provided significant survival benefits for patients with hepatocellular carcinoma [2], it is associated with significant normal tissue toxicity. It is necessary to find new effective medications for hepatocellular carcinoma. Development of pharmacologically effective agents with little toxicity or few side effects from natural products has become a new important research topic in this area. The aim of this study was to evaluate the cytotoxicity of eight triterpenoids from Thalictrum fortunei on SGC-7901 cells, Bel-7402 cell, LoVo cells and NCIH-460 cells. To study of the mechanism of cytotoxicity, cell nucleus morphology was observed using Hoechst 33258 staining, and apoptosis were analyzed by flow cytometry. To further elucidate the apoptosis pathway, the mitochondrial membrane potential (MMP), reactive oxygen species (ROS), caspase-3, P53, Bcl-2 and Bax protein expression were assayed
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