ASPP2 inhibits osteosarcoma cells metastasis by stabilizing the β-catenin-E-cadherin complex
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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
Abstract:ObjectiveTo explore the effect and mechanism of apoptosis stimulating protein2 of p53 (ASPP2) in osteosarcoma cells metastasis. MethodsQuantitative-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. ResultsThe ASPP2 mRNA levels in eight osteosarcoma tissue samples were decreased significantly than that in normal bone tissue. Compared with normal human osteoblast cells HOSB, the levels of BRMS1 mRNA and protein obviously decreased in osteosarcoma cell lines OS187、SAOS2、HOS and MG63. The metastasis ability, β-catenin-E-cadherin complex and β-catenin nuclear-translocation of SAOS2 cells was significantly inhibited after ASPP2 overexpression. ConclusionBRMS1 can inhibit SAOS2 cells metastasis and the mechanism may involve with stabilizing the β-catenin-E-cadherin complex, and suppress β-catenin nuclear-translocation. -
图 1 骨肉瘤组织及细胞中ASPP2表达的水平
A：Q-PCR骨肉瘤组织中ASPP2 mRNA表达水平; B: 细胞中ASPP2 mRNA表达水平; C：Western blot检测骨肉瘤细胞中ASPP2蛋白表达水平. *P<0.05 vs N; # P<0.05 vs HOSB.
图 2 过表达ASPP2对HOS细胞侵袭的影响
A: Western blot检测SAOS2细胞中ASPP2表达; B:Transwell实验检测SAOS2细胞侵袭能力的变化. *P<0.05 vs vector-NC.
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