| Objective:Gastric Cancer is one of the most common malignant tumors in the digestive tract. Especially in China where is a gastric cancer-prone area, the patients accounted for almost half of the total patients all over the world. Furthermore, the patients are getting younger and younger in recent years. Therefore, gastric Cancer was considered as a serious threat to human health. Jianpi Yangwei Formula (JYF), a commonly used Traditional Chinese decoction, has been proved to be an efficacious therapy for gastric cancer in clinic via the following pathways such as recurrence prevention, improvement of the symptoms, reduction of the side effects, and prolonging the survival period. However, the anticancer activity of JYF and its action mechanisms have not been fully understood. The purpose of this thesis was to explore the detailed action mechanisms of JYF in the cell and molecular biology levels and provide scientific basis for its clinical applications.Methods:1. the major active components in water and ethanol extractions of JYF were identified and simultaneously determined using liquid chromatography-mass spectrometry (LC-MS) method.2. SGC-7901stomach cancer cell lines were adopted for in vitro cytology test. Using the MTT assay, Transwell experiment and flow cytometry, the anticancer activities of JYF on SGC-7901cell lines including suppression of proliferation and invasion, arrest of cell cycle, and induction of apoptosis were fully investigated with chemical anticancer drug as positive control.3. Nude mice and SGC-7901cell lines were used to develop the nude mouse metastatic model of gastric cancer for in vivo studies. Expression changes of gastric cancer-related molecules before and after JYF extractions intervention were systematically studied by the combination use of histopathology, immunohistochemistry and Western Blotting techniques.Results:1. LC-MS analysis:several major known components of JYF were detected in both ethanol extractions and water extractions, with relatively higher content in ethanol.2. In-vitro cytology experiment:(1). Within a certain concentration range, JYF exhibited significant inhibition on cell proliferation in a dose-dependent way, with p values less than0.01compared with the control group.(2). The results of transwell experiment showed that the number of transmigrated cells in JYF group (67.3±18.3) were significantly lower than the control group (229.7±40.2) with p values less than0.01.(3). The cell apoptosis rate in JYF group was significantly higher than the control group, especially in the late apoptosis (P<0.01).(4). The G2/M period of cancer cell in JYF group was prolonged to some extent,and the S phase was shorten.3. In-vivo experiment:Compared with the control group, JYF could alleviate the weight loss of the nude mice, and significantly downregulate expression of Ki67, NMP-9, BCL-2, VEGF, PI3K, pAKT and NF-kB with p values less than0.01. Besides, JYF could activate expression of cancer resistant genes (PTEN, p<0.01) and inhibit oncogenes expression (CXCR2).Conclusion:Instead of water, ethanol is a preferred solution for JYF extraction. JYF extraction could exert various anticancer properties via the following mechanisms:inhibiting tumor cell proliferation, reducing invasion force of tumor cell, inducing tumor cell apoptosis, arresting tumor cell cycle, interfering PI3K-AKT signal transduction pathways, antiangiogenic, inhibiting oncogenes expression and inducing cancer resistant genes. Our study further proved that JYF extraction is a promising Traditional Chinese decoction for clinic therapy of gastric cancer. |
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