EBP50影响HeLa细胞微丝骨架的分布和定位

来源 :中德临床肿瘤学杂志(英文版) | 被引量 : 0次 | 上传用户:ld2001
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
Objective:To explore the influence of EBP50 (ezrin-radixin-moesin-binding phospho-protein-50) on microfila-ment 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-HA-EBP50 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. Weste 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:Weste 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. Weste 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 cytoskel-eton in cultured Hela cells and can also bind the microfilament cytoskeleton to the cell membrane under the stimulation of PDGE EBP50 may play a role in the proliferation and migration of tumor cells by influencing the distribution and localization of microfilament cytoskeleton.
其他文献
摘 要 之所以将房字加上双引号,是因为有这几种情形,一是此房处于在建阶段,二是此房已建造完毕,交付使用并办理了房屋权属证书,三是该房为存量房,也就是通常所说的“二手房”,有房屋权属证书。在第一种情形下,买卖双方还只停留在合同阶段。虽然双方签定的为《商品房买卖合同》,但并非实际上的“房屋”,合同条款注明,开发商负有在约定的时间,将具备多种条件的商品房交付买方使用,并在交付使用后通常为90日内到产权登
Objective:To study the expression of eukaryotic initiation factor-4E (eIF-4E) and p53 in squamous cell carcino-mas (SCC) and to explore their relationship and c
目的:分析乳腺癌患者健康教育中品管圈活动的应用效果.方法:抽选2014年10月至2015年10月本院收治的乳腺癌患者115例,随机分为2组,对照组57例予以常规的健康教育的方法,观察组
近日,山东省日照市五莲县一个25岁的普通女孩感动了许多人。她不幸罹患白血病,因家庭困难,主动放弃治疗,却要用生命的最后时间去帮助孩子们实现1001个微心愿。她姓郭,却不肯
摘 要 环境侵权问题不同于以往常见的侵权行为,它集中反映了侵权者与受害者之间不平等的法律关系的存在,导致受害者运用法律手段虽然做到了有法可依,却不能达到损害弥补的完美结局。因此,有必要从环境侵权最实质的问题着手,深入剖析环境侵权存在的现实问题,进而寻找最和谐的解决办法来平衡侵权者与受害者之间的关系,对环境侵权法律制度的建立和创新起到立杆见影的作用。  关键词 环境 侵权 现实问题  作者简介:周文
目的:对实践能力培养的药学实验教学体系的构建.方法:通过对实践能力培养的药学实验教学体系的建设概况进行详细的阐述,并提出一些相应的革新对策,以此来对实践能力培养的药
Objective:To detect the expression of RECK gene in the highly and low metastatic cell sublines of human os-teosarcema cell HOS and explore its possible roles on
《数学课程标准》中明确指出:要倡导积极主动、勇于探索的学习方式,强调学生的数学学习活动不应只局限于接受记忆,模仿和练习,高中数学课程还应倡导自主探索、动手实践、合作
菜单上所列的项目自然是越全越好,然而在星巴克咖啡店里,却有一款咖啡经历了数十年,仍然未能在菜单上占据一席之地,这款神秘的咖啡究竟有何奥妙呢?如果你是星巴克的常客,便会
Objective:We determine whether chemokine receptor CXCR4 and vascular endothelial growth factor (VEGF) expression related to the metastasis and survival outcome