Distribution and localization of microfilament cytoskeleton is regulated by EBP50  

EBP50影响HeLa细胞微丝骨架的分布和定位(英文)

作　　者：Kun Liu Junfang Zheng Ying Wang Peng Chen Junqi He 

机构地区：[1]Department of Biochemistry and Molecular Biology, Capital Medical University, Beijing 100069, China [2]Shandong Eye Institute, Qjngdao 266071, China

出　　处：《The Chinese-German Journal of Clinical Oncology》2009年第5期282-285,共4页中德临床肿瘤学杂志（英文版）

基　　金：Supported by grants from the National Natural Science Foundation ofChina (No. 30572183 and 30772573);Beijing Educational Committee Foundation (No. KZ200610025013);the New Century Excellent Talentsin University of China (No. NCEF-06-0184);Excellent Talents Foundation in Beijing (No. 20071D0501800253);Beijing Novagramme Foundation (No. 2008B58 [MS1]).

摘　　要：Objective:To explore the influence of EBP50(ezrin-radixin-moesin-binding phospho-protein-50) on microfilament cytoskeleton content and distribution in cultured Hela cells, and to investigate the relationship between the changes in microfilament cytoskeleton localization and EBP50 after PDGF(platelet-derived growth factor) stimulation, and to further clarify the molecular mechanism by which EBP50 suppresses tumor cell proliferation and migration.Methods:pBK-CMV-HAEBP50 wild type recombinant plasmid and pBK-CMV-HA empty vector were transfected into Hela cells.G418 at 350 mg/L was used to screen for cell clones stably expressing EBP50.Western blot was carried out to detect EBP50 expression.Similarities and differences in microfilament cytoskeleton content and distribution in Hela cells transfected with pBK-CMV-HA-EBP50 wild type recombinant plasmid and pBK-CMV-HA empty vector were also treated with PDGF(10 ng/mL and 20 ng/mL, 37 ℃, 15 min) and stained by rhodamine-labeled phalloidin to observe the distribution of microfilament cytoskeleton in the two groups.EBP50 protein distribution in PDGF-stimulated Hela cells was detected by immunofluorescence.Results:Western blot results confirmed that the EBP50 cDNA fragment could express EBP50 in cultured Hela cell lines and that cell lines stably expressing EBP50 were successfully obtained.Western blot and fluorescence results showed that in the cell line transfected with empty vector, the microfilament cytoskeleton was thick, loose, multidirectional and displayed crossing arrangements.The content of microfilament cytoskeleton in the cell line transfected with pBK-CMV-HA-EBP50 was different from that found in the cell line transfected with empty vector.EBP50 expression enhanced microfilament cytoskeleton polymerization into compact thin filaments.Under the stimulation of PDGF, EBP50 migrated to the cell membrane from the cytosol together with microfilament cytoskeleton and co-localized there.Conclusion:EBP50 can change the distribution of microfilament cytoskeleton in cult

关 键 词：microfilament cytoskeleton hela cell EBP50 

分 类 号：R318.04[医药卫生—生物医学工程] R575.2[医药卫生—基础医学]