LNMTca8113细胞中CCR10下游分子鉴定.docVIP

  LNMTca8113 细胞中 CCR10 下游分子鉴定# 张壮，潘剑* （四川大学华西口腔医院头颈肿瘤外科，成都 610041） 5 10 摘要：目的：获得与 CCR10 结合的下游分子并进行鉴定。方法：应用免疫共沉淀的方法， 获得 CCR10 共沉淀产物，通过 SDS凝胶电泳分离共沉淀产物，显示差异条带。酶切 差异条带，，蛋白酶水解，进行液相色谱-质谱联用仪检测，对共沉淀产物的肽段进行鉴定。 结果：对 CCR10 下游分子成功低进行了免疫共沉淀并有效的电泳分离，获得了差异表达条 带。液相色谱-质谱联用仪检测，20min-100min 之间有样品被洗脱。质谱图观察，峰值正常， 肽段二级质谱匹显示匹配率较高。按照筛选标准，软件检索，发现可信度高蛋白，包括 cytoskeletal 1、5、9、10 等多个分子。结论：CCR10 下游分子包括多个细胞骨架蛋白，与癌 细胞的迁移运动密切相关。CCR10 与口腔癌细胞淋巴结转移关系密切。 关键词：口腔医学；CCR10；免疫共沉淀；质谱鉴定 中图分类号：R318 15 Identification of CCR10 downstream moleculars in LNMTca8113 cells ZHANG Zhuang, PAN Jian (Department of head and neck sergery,West china stomatology hospital,Sichuan University, 20 25 30 35 40 45 ChengDu 610041) Abstract: Objective: To obtain and identify the downstream molecular binding with CCR10. Methods: CCR10 precipitation products were isolated by immunoprecipitation. Then, coprecipitation products were separated and the differential expressed bands displayed through SDSgel electrophoresis. After enzyme digestion and proteinases, liquid chromatography - mass spectrometry was used to assay and identiy peptide of coprecipitation protein. Results: CCR10 downstream molecules were successfully isolated and separated by immunoprecipitation and electrophoresis. Differentially expressed bands were further assed by liquid chromatography - mass spectrometry. A sample with normal peak of mass spectra was eluted between 20min-100min. two mass spectrometry peptide map demonstrated matching rate is higher. According to the screening criteria, the peptides with high credibility were identified by retrieval softwares, including cytoskeletal 1, 5, 9, 10, and so on. Conclusion: CCR10 downstream molecules comprise a number of cytoskeletal proteins, which is closely associated with the migration of cancer cell. Thus, CCR10 may have a important role in lymph node metastasis of oral cancer cells. Keywords: oral medicine; CCR10; immunoprecipitation; MS identification 0 引言 炎症趋化因子是一个多肽信号分子家族，通过激活 G 蛋白偶联受体，参与细胞趋化、 白细胞的归巢、细胞