胃癌细胞多药耐药相关蛋白质筛选鉴定的蛋白质组学研究

Identification of multidrug resistance related proteins in different gastric carcinoma cell lines by comparative proteomics

目的在胃癌细胞株SGC7901及稳定的耐药细胞株SGC7901／VCR中寻找与胃癌多药耐药（MDR）直接相关的蛋白质，观察其功能。方法胃癌细胞株SGC7901和稳定的耐药细胞株SGC7901／VCR培养后提取总蛋白，采用蛋白质组学技术（双向凝胶电泳和基质辅助激光解吸电离-飞行时间质谱鉴定）筛选并鉴定差异表达的蛋白质，并应用蛋白印迹试验法对鉴定出的部分蛋白质在胃癌细胞株中的表达进行验证。结果在两种细胞株中找到表达差异明显的蛋白质点9个，经质谱分析，7种蛋白质得到鉴定，为S60核糖体蛋白L23、电压依赖性阴离子选择性通道蛋白1、锌指蛋白394、角蛋白I型细胞骨架9、核糖核酸3’-端磷酸化酶1、matrin型锌指蛋白2、Sideroflexin-1。这些蛋白质与胃癌MDR直接相关。蛋白印迹试验法检测部分蛋白质在胃癌细胞株中的表达，与蛋白质组学所得结果一致。结论胃癌细胞SGC7901和SGC7901／VCR蛋白质表达存在差异。

Objective To investigate multidrug resistance related proteins in human gastric carcinoma cell lines SGC7901 and SGC7901/VCR by comparative proteomics. Methods After culture, the holoproteins of human gastric carcinoma cell lines SGC7901 and GC7901/VCR were extracted, and then measured by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization time-of- flight mass spectrometry （MALDI-TOF-MS）. Some proteins obtained through proteomics were tested by Western blotting in the cell lines. Results There were different proteins in these 2 human gastric carcino- ma cell lines. Nine different protein spots were found in these 2 gastric carcinoma cell lines, and 7 spots were identified by MALDI-TOF-MS, and these proteins were 60S ribosomal protein L23 （RL23）, voltage- dependent anion-selective channel protein-1 （ VDAC1 ）, zinc finger protein （ZNF） 394, keratin, type I cytoskeletal 9 （KICg）, RNA 3＇ -terminal phosphate cyclase 1 （RTC1）, zinc finger matrin-type protein 2, Sideroflexin-1. These proteins had a direct relationship with multidrug resistance in gastric carcinoma. Results of Western blotting about protein expression were in consonance with those of proteomics. Conclusion Different proteins were found in human gastric carcinoma cell lines SGC7901 and SGC7901/VCR.