Natural products certainly are a precious source of promising leads for the development of novel cancer therapeutics. proliferation and metastasis of MDA-MB-231 cells. It could be a promising agent for breast cancer therapy. (Sam.) Juzep, an aquatic plant, belonging to the Alismataceae family, which A 83-01 can be distributed in China broadly, Korea, and Japan [7]. In China, A 83-01 it’s been broadly utilized like a folk hypolipidemic and diuretic real estate agents for greater than a thousand years, and continues to be used for the treating dysuria, hypertension, edema, and urinary system attacks [7,8,9]. Contemporary pharmacological investigations possess proven the diuretic, anti-hypertensive, anti-cancer, hypoglycemic, and anti-atherosclerotic activities of Alismatis Rhizoma [7,10,11,12,13,14,15]. The chemical constituents of Alismatis rhizome mainly consist of triterpenoids, polysaccharides, sesquiterpenes, diterpenes, and essential oil [16]. Alisol A (Figure 1A), belonging to protostane-type tetracyclic triterpenoid, serves as one of the main components in Alismatis Rhizoma. However, there is little information concerning its anti-cancer activity. In this study, we investigated the anti-cancer activity A 83-01 of alisol A in human breast cancer cells and attempted to elucidate its possible molecular mechanism. Open in a separate window Figure 1 Effects of alisol A on cell viability in human breast cancer cells. (A) The chemical structure of alisol A. (B) Effects of alisol A on cell viability in MDA-MB-231, MDA-MB-453, and MCF-7 cells. Cells were treated with different concentrations of alisol A for 24 h. Then, cell viability was quantified by the MTT assay. Data represent the mean S.D of at least three independent experiments. * < 0.05, ** < 0.01, *** < 0.001. 2. Material and Methods 2.1. Cell Culture A 83-01 and Reagents MDA-MB-231, MCF-7, and MDA-MB-453 cell lines were purchased from the Cell Rabbit Polyclonal to CYSLTR1 Bank of the Institute of Biochemistry and Cell Biology, China Academy of Sciences (Shanghai, China) and stored in liquid nitrogen. Cells were cultured in DMEM culture medium (Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS, Gibco, USA), 100 U/mL penicillin G, 2.5 g/mL amphotericin B, and 100 g/mL streptomycin (complete medium) at 37 C with 5% CO2 in a humidified atmosphere. Alisol A was purchased from MedChemExpress (Monmouth Junction, NJ, USA) (The chemical structure is shown in Figure 1A). 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2< 0.05; ** < 0.01; *** < 0.001. 3. Results 3.1. Effects of Alisol A on Cell Viability in Human Breast Cancer Cells To determine the effects of alisol A on the growth of human breast cancer cells, the cytotoxic effects were measured by MTT assay. Breast cancer is a heterogeneous disease with high degree of diversity based on histology, cellular origin, metastatic potential, therapeutic response, and clinical outcome [17]. Generally, there are three identified types: HER2 (+), ER/PR (+), and TNBC (defined by the lack of ER, PR, and HER2 in breast cancer cells) breast cancer cells [18]. In the present study, MDA-MB-231 (TNBC), MCF-7 (ER/PR (+)) and MDA-MB-453 (HER2 (+)) cell lines were used. Cells were treated with different concentrations of alisol A for 24 h. As shown in Figure 1B, alisol A significantly inhibited the growth of MDA-MB-231 cells in a concentration-dependent manner. However, alisol A did not show obvious cytotoxic effects on MCF-7 and MDA-MB-453 cells. Therefore, MDA-MB-231 cells were considered as an in vitro model for further study. 3.2. Effects of Alisol A on Induction of Cell Apoptosis To determine whether the growth inhibitory effects of alisol A were associated with A 83-01 the induction of apoptosis, Annexin V-FITC/PI double staining was used as a criterion to distinguish apoptotic cells by flow cytometry analysis. As shown in Figure 2A, alisol A treatment for 24 h did not boost the amount of apoptotic cells in MDA-MB-231 cells significantly. The percentage of apoptotic cells was improved from 9.90 .