菝葜提取物抗卵巢癌的多重效应及其分子机制-药剂学专业论文.docxVIP

华中科技大学硕士学位论文 华 中 科 技 大 学 硕 士 学 位 论 文 PAGE 2 PAGE 2 ABSTRACT Objective Smilax china L. rhizome (SCR) has been proposed to act as an anti-inflammatory and chemopreventive agent. In the present study, the antitumor effects of the SCR extracts on ovarian cancer cells and the mechanisms involved were investigated. Methods Ovarian cancer cells were treated with SCR extracts. Firstly, the effect on cell viability， proliferation and colony capability were evaluated by CCK-8 assay, EdU incorporation assay and colony assay, respectively. And the sensitization of ovarian cancer cell A2780 to cisplatin and adriamycin by SCR extracts were also analyzed by CCK-8 assay. Secondly, Cells stained by propidium iodide and Annexin V-FITC/PI, then analyzed by flow cytometry for cell cycle and apoptosis, respectively. Thirdly, Transwell were used to determine cell migration. Finally, several proteins were determined by western blotting, and cellular distribution of NF-κB was observed by immunofluorescence. Results SCR extracts from BuOH (SCR-B) suppressed the viability, proliferation and colony capability of A2780 cells in a dose-dependent manner. Furthermore, we discovered that SCR-B promoted chemo-sensitivity to cisplatin and adriamycin in A2780 cells. Both Transwell and wound-healing assay demonstrated that SCR-B attenuated migration of A2780 cells. Treatment of SCR-B resulted in induced G2/M arrest and induced apoptosis in A2780 cells through activation of Bax , caspase-3 and PARP, and the inhibition of NF-κB, which resulted to down-regulation of Bcl-2, Bcl-xL, XIAP, and cIAP-1 via the signaling pathway. AKT, one of NF-κB’s important upstream elements, and the NF-κB -regulated gene products VEGF, ICAM-1 and Cyclin D1 were found to be down-regulated by SCR-B. Conclusions Our results suggest that SCR-B is an effective anti-proliferation agent of ovarian cancer cells, the effect is might caused by inhibition of NF-κB, it also found that SCR-B can reduce chemo-resistance. The results fr