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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >SPECIFIC SAMPLE PREPARATION IN COLORECTAL CANCER
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SPECIFIC SAMPLE PREPARATION IN COLORECTAL CANCER

机译：大肠癌的特殊样品制备

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Large tissue samples from ten patients operated for colorectal cancer were prepared in the operating roam in iced phosphate buffered saline, containing ethylene diaminetetraacetic acid and protease inhibitors. After cutting the specimens into small fragments, the tissues were gently pressed through a steel mesh. Membranes were permeabilized in chilled ethanol 70% to allow cytosolic fluoresceine isothiocyanate labeling, pet-formed with anti-cytokeratin (CAM 5.2) antibodies. Samples were quantitatively sorted with a fluorescence activated cell sorter (FAGS) and denatured before processing separation by two-dimensional electrophoresis on polyacrylamide gels, This Procedure made it possible to sample about 4 x 10(7) viable normal and tumoral cells before fixation, and up to 4 x 10(6) cells after FAGS, The gels run before and after fixation showed no major differences. The rate of cytokeratin-positive cells in the samples was the following (mean, Cl 5-95%): mucosa 29.5% (8.9-66.7%), tumor 44.3% (6.6-94.8%). The epithelial cell content in colorectal cancer and normal mucosa shows important intersample variations. This is important for any comparison of fresh samples, whether at DNA, RNA, or at the protein level, We propose a method allowing the preparation of pure epithelial cell samples from normal and tumoral colonic fresh mucosa. [References: 10]
机译：在冰冻的磷酸盐缓冲液中漫游，准备了十名接受结肠直肠癌手术的患者的大组织样本，其中含有乙二胺四乙酸和蛋白酶抑制剂。将标本切成小块后，将组织轻轻地压过钢丝网。将膜在70％的冷乙醇中通透，以胞质标记异硫氰酸荧光素异硫氰酸酯，并用抗细胞角蛋白（CAM 5.2）抗体形成宠物。样品通过荧光激活细胞分选仪（FAGS）进行定量分选，并变性，然后在聚丙烯酰胺凝胶上进行二维电泳分离，从而进行分离。该程序使得在固定之前可以对大约4 x 10（7）正常和肿瘤细胞进行采样，并且FAGS后最多4 x 10（6）个细胞，在固定之前和之后运行的凝胶均无重大差异。样品中细胞角蛋白阳性细胞的比率如下（平均值，Cl 5-95％）：粘膜29.5％（8.9-66.7％），肿瘤44.3％（6.6-94.8％）。大肠癌和正常粘膜中的上皮细胞含量显示出重要的样本间差异。这对于无论是DNA，RNA还是蛋白质水平的新鲜样品的任何比较都是重要的。我们提出了一种允许从正常和肿瘤结肠新鲜粘膜制备纯上皮细胞样品的方法。 [参考：10]

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