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目的建立高分辨率的移行上皮癌细胞PUMC双向凝胶电泳图谱,并初步分析其蛋白质表达情况。方法用固相pH梯度双向凝胶电泳技术分离膀胱移行上皮癌细胞PUMC细胞总蛋白,凝胶考马斯亮蓝染色,然后利用GS-800 Calibrated Densitometer凝胶成像系统获取图像,用 PDQuest2D分析软件对图像进行背景消减、斑点检测、匹配、斑点位置重复性进行系统分析。结果获得了背景清晰、分辨率和重复性较好的双向凝胶电泳图谱。结论建立膀胱癌移行上皮癌细胞PUMC的双向电泳参考图谱,为进一步构建其蛋白质表达数据库提供了基础。
OBJECTIVE: To establish a high-resolution PUMC two-dimensional gel electrophoresis (PUMC) pattern of transitional cell carcinoma of the epithelium and to analyze its protein expression. Methods The total protein of PUMC cells in transitional cell carcinoma of urinary bladder was separated by two-dimensional gel electrophoresis with solid-phase pH gradient. Coomassie brilliant blue staining was performed on the bladder transitional epithelial carcinoma cells. Then the images were obtained by using the GS-800 Calibrated Densitometer gel imaging system and PDQuest2D software Background subtraction, speckle detection, matching, and spot location repeatability were systematically analyzed. Results Two-dimensional gel electrophoresis with clear background, good resolution and repeatability was obtained. Conclusion The establishment of two-dimensional electrophoresis reference map of PUMC in transitional cell carcinoma of the bladder cancer provided the basis for further construction of its protein expression database.