人食管癌Eca-109细胞膜蛋白的分离和纯化

Isolation and purification of membrane proteins of human esophageal carcinoma Eca-109 cell line

目的:建立人食管癌Eca109细胞系膜蛋白分离和初步纯化方法。方法:使用改良的Neville法提取细胞膜,在pH6.3的条件下,用去垢剂TritonX100和辛基β葡糖苷把细胞膜上的蛋白溶解下来,以超滤法纯化膜蛋白并分组。结果:在pH6.3的条件下,适当的去垢剂与膜蛋白质量之比使用去垢剂辛基β葡糖苷时为61;使用TritonX100时为21。超滤法将膜蛋白分为相对分子质量<3000、3000～10000、<10000、10000～30000、30000～100000、>100000共6个组分。结论:获得适当的去垢剂与膜蛋白质量比,并获得了不同的膜蛋白组分,为进一步研究人食管癌Eca109细胞肿瘤抗原奠定了基础。

Aim ： To establish the initial methods to isolate and purify the membrane proteins of human esophageal carcinoma Eca-109 cell line. Methods：The cell membrane was extracted with the improved Neville＇s method（ pH 6.3）. The membrane proteins were dissolved out from cytomembrane with the detergents Triton X-100 or Octyl-β-D-glucopyranoside, respectively. Results： The appropriate ratio of detergent （Octyl-β-D-glucopyranoside） to protein was 6 ： 1. In the Triton X-100 group the ratio was 2 ： 1. The membrane proteins were allocated into 6 components including Mr 〈 3 000, 3 000 N 10 000, 〈 10 000, 10 000 N30 000, 30 000 - 100 000, 〉 100 000. Conclusion： This study provides foundation to investigate the tumor antigen of human esophageal carcinoma Eca-109 cell line.