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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。
OBJECTIVE: To analyze the effect of total RNA of Pinellia tuber on the protein expression profiles of human ovarian cancer SKOV3 cells. Methods: The CCK-8 method was used to determine the concentration of total protein in ovary cancer SKOV3 cells treated with Pinellia tuber. The total protein of total RNA extracted from Pinellia ternata and untreated SKOV3 cells was separated by two-dimensional gel electrophoresis (2-DE) using immobilized pH gradient (IPG) PDQuest7.4.0 comparative analysis of two-way gel electrophoresis of the palm leaf Pinellia total protein-treated group and untreated group SKOV3 cell lines and identify differentially expressed proteins. Results: The two-dimensional gel electrophoresis pattern of SKOV3 cell line with clear background, high resolution and good reproducibility was obtained. The protein spots of untreated and untreated group were 964 ± 6 and 883 ± 20, respectively. The average match rate Respectively: 99% and 96%. Pinellia total protein treatment group of three plastic in the protein spots have good repeatability. Two hundred and seventy five protein spots were screened out from the two-dimensional gel electrophoresis of SKOV3 cells treated with total protein of Pinellia tuber and untreated group, of which 154 were up-regulated, 178 down-regulated, 96 were expressed only in untreated group, 122 Only in the palm leaf Pinellia total protein treatment group expression. CONCLUSION: Two-dimensional gel electrophoresis pattern of total tuber treated with untreated SKOV3 cells was established for the first time, and 550 differentially expressed protein spots were identified. The identification of these differentially expressed proteins will reveal the anti-ovarian cancer effect The mechanism to provide experimental evidence.