LNMTca8113舌癌细胞中CCR10下游分子鉴定  

Identification of CCR10 downstream moleculars in LNMTca8113 tongue cancer cells

作　　者：王卓敏[1] 李迎春[1] 张壮[2] 

机构地区：[1]解放军第452医院核医学科,四川成都610021 [2]四川大学华西口腔医院头颈肿瘤外科,四川成都610041

出　　处：《现代肿瘤医学》2013年第12期2644-2647,共4页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:30901690);教育部博士点新教师基金资助项目(编号:200901811200 36)

摘　　要：目的:在LNMTca8113舌癌细胞中获得与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LNMTca8113舌癌细胞中CCR10的共沉淀产物,通过SDS-PAGE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下游分子成功地进行了免疫共沉淀及有效的电泳分离,获得了差异表达条带。液相色谱-质谱(LC-MS)联用仪检测,20min-100min之间有样品被洗脱。质谱图观察,峰值正常,肽段二级质谱匹配图显示匹配率较高。按照筛选标准,通过软件检索,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与舌癌细胞淋巴结转移关系密切。Objective：To obtain and identify the downstream moleculars binding with CCR10. Methods：CCR10 precipitation products in LNMTca8113 tongue cancer cells were isolated by immune - precipitation. Then, co - precip- itation products were separated and the differential expressed bands displayed through SDS -PAGE gel electrophore- sis. After enzyme digestion and proteolysis,liquid chromatography - mass spectrometry was used to assay and identify peptide of co - precipitation protein. Results： CCR10 downstream molecules were successfully isolated and separated by immune - precipitation and electrophoresis. Differentially expressed bands were further asseessd by liquid chroma- tography -mass spectrometry （LC - MS）. A sample with normal peak of mass spectra was eluted between 20min - 100min. Two mass spectrometry peptide map demonstrated matching rate was higher. According to the screening crite- ria,the peptides with high credibility were identified by retrieval softwares, including cytoskeletal 1,5,9,10, and so on. Conclusion：CCRl0 downstream molecules comprise of a number of cytoskeletal proteins, which are closely associ- ated with the migration of cancer cells. Thus, CCR10 may have an important role in lymph node metastasis of tongue cancer cells.

关 键 词：舌癌 淋巴结转移 CCR10 免疫共沉淀 质谱鉴定 

分 类 号：R730.43[医药卫生—肿瘤] R739.86[医药卫生—临床医学]