应用双向凝胶电泳研究掌叶半夏总蛋白对人卵巢癌SKOV3细胞蛋白质表达谱影响.docVIP

应用双向凝胶电泳研究掌叶半夏总蛋白对人卵巢癌SKOV3细胞蛋白质表达谱影响摘 要:目的:分析掌叶半夏总蛋白对人卵巢癌SKOV3细胞蛋白质表达谱的影响。方法:CCK-8法分析确立掌叶半夏总蛋白处理卵巢癌SKOV3细胞的浓度。应用固相 pH梯度 ( immobilized pH gradient, IPG)双向凝胶电泳技术 ( two-dimensional gel electrophoresis, 2-DE)分离掌叶半夏总蛋白处理组和未处理组SKOV3细胞的总蛋白,银染显色,PDQuest7.4.0比较分析掌叶半夏总蛋白处理组和未处理组SKOV3细胞系的双向凝胶电泳图谱并识别差异表达的蛋白质。结果:获得了背景清晰、分辨率高、重复性好的SKOV3细胞系的双向凝胶电泳图谱,掌叶半夏总蛋白未处理组与处理组蛋白质点数为:964±6和883±20;平均匹配率分别为:99%和 96%。掌叶半夏总蛋白处理组的 3块胶在蛋白质点位置上有较好的重复性。比较分析掌叶半夏总蛋白处理组和未处理组SKOV3细胞的双向凝胶电泳图谱,筛选出差异蛋白质点550个,其中154个表达上调,178个表达下调,96个仅在未处理组表达,122个仅在掌叶半夏总蛋白处理组表达。结论:首次建立了掌叶半夏总蛋白处理和未处理SKOV3细胞的双向凝胶电泳图谱,识别了550个差异表达的蛋白质点,对这些差异蛋白质点的鉴定将为揭示掌叶半夏总蛋白抗卵巢癌作用的机理提供实验依据。 关键词:掌叶半夏总蛋白;卵巢癌;双向凝胶电泳;蛋白质组? 中图分类号:R737.31 文献标识码:A 文章编号:1673-7717(2010)04-0789-04 [FQ(4。52,X-W] 收稿日期:2009-10-28 基金项目:浙江省中医药科研项目(2006Y043) 作者简介:周莉(1970-),女,副教授,博士,研究方向:妇科肿瘤。 Alteration of Protein Profiles in Human Ovarian Cancer Cell Line SKOV3 Treated with Protein of Pinellia Pedatisecta Schott. ZHOU Li,WANG Han?chu, ZHENG Fei?yun, SHI Meng?ru (Department of Gynecology and Obstetrics, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang,China) Abstract:?Objective:?To evaluate the effect of the Protein ofPinellia pedatisecta Schott treated human ovarian cancer cell line SKOV3 on protein expression. Methods: CCK-8 assay was used to determine the concentration of the Protein ofPinellia pedatisecta Schott in treating SKOV3. The total proteins of control and the Protein of Pinellia pedatisecta Schott-treated SKOV3 cells were separated with immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). After silver staining, the gel images were acquired by image scanner and then analyzed with the PDQuest7.4.0 software. 2-DE maps of control and the Protein of Pinellia pedatisecta Schott-treated SKOV3 cells were established. ?Results:? The IC50 of the Protein of Pinellia pedatisecta Schott exposed SKOV3 cells for 72 hours was 82.396 μg/mL. Well-resolved, reproducible 2-DE patterns of control and