肉细胞转移.PDF

·40 · doi 10.3969/j.issn. 1674-4500.2017.01.12 分子影像学分子影像第年第4040卷第卷第11期期 临床研究 pp5353 凋凋亡刺激蛋白亡刺激蛋白22 通过稳定通过稳定ββ-catenin-E-cadherin-catenin-E-cadherin 复合体抑制骨复合体抑制骨 肉细胞转移肉细胞转移 郑少伟，钟浩博，缪海雄，刘伟乐，李胜发，陈楚群，孙春汉 惠州市第一人民医院骨科，广东 惠州 516001 摘要：目的探讨p53 凋亡刺激蛋白2 （ASPP2）对骨肉瘤细胞转移的影响及其机制。方法 Q-PCR 检测骨肉瘤组织和细胞中 ASPP2 mRNA 的表达；western blot 检测骨肉瘤细胞中ASPP2 蛋白表达；transwell 检测细胞转移能力；co-IP 实验检测β-catenin 和E-cadherin 相互作用水平；免疫荧光检测β-catenin在细胞核中表达水平。结果 8例手术标本Q-PCR 检测发现，骨肉瘤组织中 ASPP2 mRNA 的表达较癌旁组织明显降低；4 株骨肉瘤细胞系中（OS187、SAOS2、HOS 和MG63）ASPP2 mRNA 和蛋白的表达 较正常成骨细胞明显减少。过表达ASPP2 蛋白的SAOS2 其转移能力明显减弱，同时β-catenin 和E-cadherin 相互作用水平增 加，而β-catenin 核转位减少。结论 ASPP2 可以减少骨肉瘤细胞SAOS2 的转移，其机制可能的通过稳定β-catenin-E-cadherin 复 合体，抑制β-catenin 核转位。 关键词：p53凋亡刺激蛋白2 ；骨肉瘤转移；β-catenin-E-cadherin 复合体；β-catenin 核转位 ASPP2 inhibits osteosarcoma cells metastasis by stabilizing the β-catenin-E-cadherin complex ZHENG Shaowei, ZHONG Haobo, MIAO Haixiong, LIU Weile, LI Shengfa ,CHEN Chuqun, SUN Chunhan Department of Orthopedic Surgery, Huizhou First Peoples Hospital, Huizhou 516001, China Abstract: Objective To explore the effect and mechanism of apoptosis stimulating protein2 of p53 (ASPP2) in osteosarcoma cells metastasis. Methods Quantitative-Polymerase Chain Reaction (Q-PCR) was performed to determine ASPP2 mRNA in eight samples of osteosarcoma tissue and four cell lines of osteosarcoma. The levels of ASPP2 protein were detected by western blot. The metastasis ability of SAOS2 cells that overexpressed ASPP2 were estimated by Transwell experiment. The Levels of interaction of β-catenin and E-cadherin were analyze by co-IP. Immunofluorescence was used to detect the expression of ASPP2 protein in the nuclear of osteosarcoma cells. Results The ASPP2 mRNA levels in eight osteosarcoma tissue samples were decreased significantly than that in n