| Ovarian carcinoma is one of the most common malignant gynecological tumors. Owing to its rapid growth and early abdominal dissemination, the long-term survival of patients is rather low, usually under 30%. Therefore, it is of great significance to develop more effective intraabdominal chemotherapy. Recently, some ingredient extracted from herbal medicine and natural plant is becoming new approaches to prevent and cure tumor. Genistein is one of the main ingredients of isoflavones, widely distributed in many dietary plants such as soybeans, memdranous milkvetch root and loded kudzurine root, with many physiological and pharmacological activities. It is regarded commonly that Genstein is a potential tumor inhibitor. Although it has studied in various tumors such as leukaemia, breast carcinoma, prostate carcinoma, lung cancer, carcinoma of the large intestine, lymphoma and melanoma, there are few reports regarding its effect and mechanism in ovarian malignant tumors, especially the most common clinical type-ovarian serous cystadenocarcinoma. In this study, light and electron microscopy, MTT, AO/EB fluorescence staining, flow cytometry, immunohistochemistry, RT-PCR, in situ hybridization, Western blotting were used to explore the effects and molecular mechanisms of Genistein on growth and invasion inhibition in human ovarian carcinoma cell line SKOVs and its xenograft in nude mice. The study is divided into four parts: (1) Effect of Genistein on growth inhibition in SKOV3 cell and its xenograft in nude mice; (2) Molecular mechanisms of Genistein on proliferation inhibition and apoptosis induction in SKOV3 and its xenograft in nude mice; (3) Effect of Genistein on invasion inhibition in SKOV3 cell and its xenograft in nude mice; (4) Molecular mechanisms of Genistein on invasion inhibition in SKOV3 cell and its xenograft in nude mice.The aim is to provide experimental data for prevention and therapy of ovarian carcinoma and clinical application of Genistein.Themain results are as follows:1. SKOV3 cells were treated with 5-40 Mmol/L Genistein for 24 to 72 hours or treated with 20Pmol/L Genistein for 72 hours to investigate the effect of growth inhibition and apoptosis inducement. Antiproliferative effect of Genistein against SKOV3 was tested by MTT method. A time-dependent and dose-dependent growth inhibition was demonstrated. The characteristic apoptotic morphological changes were observed in SKOV3 cells after treated with Genistein by AO/EB fluorescence staining and electron microscope observation. Flow cytometric analysis revealed that 20 mol/L Genistein induced a sub-G1 peak and a G2/M phase arrest. It was demonstrated that Genistein (0.4mg/kg, s.c.) inhibited the transplanted xenograft growth in vivo in this study. The main finding included that the tumor volume and tumor weight decreased, T/C ratio (mean volume of treated group/mean volume of control group) also was reduced compared to untreated group, and extensive necrotic areas in the tumor treated with Genistein appeared. A weight loss of nude mice after treatment with Genistein was not significant as compared with the control group. All these results suggested that PTK inhibitor Genistein is effective in the treatment of cell line SKOV3 without obvious toxicity. And when treated with Genistein (0.4mg/kg, s.c.) and Cisplatin (4mg/kg, i.v.), it showed a synergistic effect on human ovarian cancer cell line SKOV3 and its xenograft in nude mice.2. In this study, the expression of proliferation and apoptosis associated proteins and their mRNA were detected using immunocytochemistry, RT-PCR and Western blotting in SKOV3 and its xenograft treated with Genistein. The expression of PCNA and CyclinB1 protein were decreased, p2iWAF1/CIP1 protein and mRNA were increased. Genistein interrupted cell cycle, induced a G2/M cell cycle arrest and inhibited cell proliferation possibly due to the down-regulation of PCNA and cyclinB1, up-regulation of p21WAF1/CIP1. Genistein also down-regulate the expression of c-raf-1 mRNA, C-erbB2 and c-jun and c-fos protein and...
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