乙醇固定对肿瘤细胞的蛋白质提取及Western印迹的影响

为研究乙醇固定后肿瘤细胞的蛋白质提取及Western印迹的可行性。于低温（4℃）或常温条件下裂解经75％冷乙醇固定的肿瘤细胞（HL－60细胞和Molt-4细胞），提取细胞蛋白质进行SDS－PAGE凝胶电泳和Western印迹。结果显示经预处理后的肿瘤细胞中提取的蛋白质不论是在SDS－PAGE后的考马斯亮蓝染色，还是转膜后的Western印迹均可得到可分析的蛋白质条带。提示75％冷乙醇固定的肿瘤细胞仍适用于SDS-PAGE和Western印迹。并可结合流式细胞术对肿瘤细胞蛋白质进行细胞周期定性、定量和定时相的同步分析。

Effect of Ethanol Fixation on Protein Extraction and Western Blotting in Tumor Cells

The feasibility of protein extraction and Western blotting from ethanol-fixed tumor cells was studied. Cold ethanol (75 %) fixed tumor cells (HL-60 and Molt-4) were lysised and protein extracted under room temperature or hypothermia. Then sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of protein with subsequent Western blotting was performed. It was found that protein bands from cold ethanol-fixed tumor cells on the polyacrylamide gel with coomassie brilliant blue stain or nitrocellulose filter with Western blotting could be used for comparative analysis. The results demonstrated that cold ethanol fixation could prevent tumor cells proteins from degrading. These tumor cells can be used for Western blotting, and by combined with flow cytometry can be used to analyze the position and quantity of the target protein expression in cell cycle.