中国肿瘤生物治疗杂志2018，Vol.25Issue(3)：258-262,5.DOI:10.3872/j.issn.1007-385x.2018.03.008
宫颈癌HeLa细胞内SKP2结合蛋白的筛选和功能预测
Screening of S-phase kinase-assuciated protein 2 in the cervical carcinoma Hela cell
摘要
Abstract
Objective:The co-immunoprecipitation and mass spectrometric analysis was carried out to obtain the S-phase kinase-associated protein 2 (SKP2)-binding proteins in HeLa cells,and the biological functions of these binding proteins were forecast.Methods:The co-immunoprecipitation system was established by co-immunoprecipitation and Western blotting assay;the specific protein gel of SKP2-binding proteins was obtained by SDS-PAGE and silver staining assay;the potential SKP2-binding proteins was identified by mass spectrometric analysis;and the GO (Gene ontology) analysis and KEGG analysis was carried out by bioinformatics technique.Results:The expression level of SKP2 protein in HeLa cells was high enough for co-immunoprecipitation assay;the co-immunoprecipitation system was established successfully,and SKP2-binding proteins was obtained;a total of 563 proteins were identified by mass spectrometric analysis,and 270 proteins with high credibility were obtained after screening.The GO analysis and KEGG analysis was carried out for the 270 proteins to forecast their functions and pathways.Conclusion:The SKP2-binding proteins were screened successfully,and it was the foundation for the subsequent screening of target-binding proteins and the search for targeting drugs.关键词
细胞S期激酶相关蛋白2/免疫共沉淀/宫颈癌/结合蛋白Key words
S-phase kinase-associated protein 2/co-immunoprecipitation/cervical cancer/binding-proteins分类
医药卫生引用本文复制引用
贾静,方健飞,任娟,贾振宇,王孝举..宫颈癌HeLa细胞内SKP2结合蛋白的筛选和功能预测[J].中国肿瘤生物治疗杂志,2018,25(3):258-262,5.基金项目
浙江省自然科学基金资助项目(No.LY18H160067,LQ12H16013) （No.LY18H160067,LQ12H16013）
浙江省医药卫生科技计划资助项目(No.2015KYB090,2011KYB005). Project supported by the Natural Science Foundation of Zhejiang Province(No.LY18H160067,LQ12H16013),and the Zhejiang Provincial Medical and Healthy Foundation for Science and Technology Program(No.2015KYB090,2011KYB005) （No.2015KYB090,2011KYB005）
