All experiments were performed three times and expressed as mean SD. belongs to the composite family has been an important traditional herbal medicine in Asia, which is usually widely used to treat dyspepsia, diarrhea, stomach diseases, diabetes and anti-abortion [9,10,11,12]. It is also popularly used as heath cultivating food. There are a large number of natural compounds that extracted from shows a wide range of biological and pharmacological activities, for example, against insomnia and anxiety, neuroprotective, platelet activation and anti-cancer effect [14,15,16,17]. Previous studies reported that AT-II could inhibit cell proliferation, arrest G1 phase cell cycle and induce apoptosis in B16 cell . However, the effects and mechanisms of AT-II on human gastric cancer remain elusive. The purpose of our study is to investigate the effects of AT-II on cell proliferation, motility and apoptosis of gastric carcinoma cells Diosbulbin B and its possible molecular mechanisms, which would provide valid data for the Rabbit Polyclonal to p38 MAPK (phospho-Thr179+Tyr181) application of AT-II to treat Diosbulbin B gastric carcinoma in the future. 2. Results 2.1. AT-II Inhibits Proliferation in HGC-27 and AGS Cells To research the effects of AT-II on cell growth, CCK-8 assays were used to determine relative cell viability. As shown in Physique 1, AT-II treatment groups showed significant inhibitory effects on HGC-27 and AGS cells compared to control group in a concentration and time-dependent manner. Moreover, HGC-27 cells are more sensitive than AGS cells to AT-II. When HGC-27 cells were exposed to 200 M of AT-II for 48 h, the cell viability reduced to nearly 50%, while AGS needed 400 M of AT-II treatment. However, even if treated with 400 M of AT-II for 48 h, it had no cytotoxicity on human normal gastric mucosal epithelium GES-1 cells. Open in a separate window Physique 1 Inhibitory effects of AT-II on cancer cell growth. (A) HGC-27, (B) AGS and (C) GES-1 cells were treated with AT-II at different concentrations for 24 h, 48 h and 72 h. Cell viability was examined by CCK-8 assay. All data were obtained from three impartial experiments and expressed as mean SD. * < 0.05 and ** < 0.01 vs. control group. 2.2. AT-II Affects Morphological Changes After being treated with AT-II for 48 h, the morphological changes of HGC-27 and AGS cells were observed with an inverted microscope, which had remarkable differences from the control group. Compared to control group, the majority of HGC-27 and AGS cells treated with AT-II were obviously reduced, distorted and grew Diosbulbin B slowly. In addition, with a high dose of AT-II treatment, the cell membrane became rough and emerged blebbing and swelling (Physique 2). Open in a separate window Physique 2 Morphological changes of HGC-27 and AGS cells treated with AT-II for 48 h and observed with an inverted microscope 200 magnification. (A) HGC-27 cells; (B) AGS cells. 2.3. AT-II Induces Apoptosis in HGC-27 and AGS Cells HGC-27 and AGS cells were treated with various doses of AT-II for 48 h and stained with Annexin V-FITC/Propidium Iodide (PI). Flow cytometry results exhibited that cell apoptosis rates of HGC-27 and AGS cells were positively correlated with the concentration of AT-II. The upper right quadrant represented late apoptotic cells and the lower right quadrant represented early apoptotic cells. Treated with 50 M of AT-II, cell apoptosis rate did not have differences from control group in HGC-27 cells, but the percentages of apoptotic cells were significantly increased with the increasing AT-II concentrations (Physique 3A). However, AGS cells were less sensitive to AT-II than HGC-27 cells and when only exposed to 200 M of AT-II, AGS cells could exhibit remarkable apoptosis (Physique 3B). Open in a separate window Physique 3 Apoptotic effects of AT-II on HGC-27 and AGS cells for 48 h. (A) HGC-27 cells; (B) AGS cells; (C) the percentages of apoptotic cells in HGC-27 cells; (D) the percentages of apoptotic cells in AGS cells. All experiments were performed in triplicates and expressed as mean SD. * < 0.05 vs. control group. 2.4. AT-II Suppresses the Capability of Cell Motility To determine the effect of AT-II on cell.