| 罗玮周天鸿闫道广.ORP8 与SPAG5 相互作用并影响细胞周期[J].现代生物医学进展英文版,2011,11(14):2601-2604. |
| ORP8 与SPAG5 相互作用并影响细胞周期 |
| Interaction of ORP8 with SPAG5 and Their Effects on Cell Cycle |
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| DOI： |
| 中文关键词: ORP8 酵母双杂交 SPAG5 蛋白质相互作用 细胞周期 |
| 英文关键词: ORP8 Yeast two hybrid SPAG5 Protein interaction Cell cycle |
| 基金项目:国家自然科学基金（30971104） |
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| 中文摘要: |
| 目的：筛选与氧化固醇结合蛋白相关蛋白8 （Oyxterol binding protein related protein 8，ORP8） 相互作用的蛋白质，为揭示
ORP8 在细胞中的功能及其机制提供线索，并根据相互作用蛋白质初步探索ORP8 的功能。方法：构建重组诱饵质粒
pGBKT7-ORP8m，利用GAL4 酵母双杂交系统筛选人Universal cDNA 文库，筛选出与ORP8 相互作用的蛋白质，通过GST
Pull-down 以及免疫共沉淀（Co-immunoprecipitation，Co-IP）验证蛋白质相互作用，并通过流式细胞仪检测过表达ORP8 对HepG2
细胞周期的影响。结果：酵母双杂交筛选得到了精子相关抗原5 （Homo sapiens sperm associated antigen 5，SPAG5），体外GST
pull-down 和Co-IP 实验确证了ORP8-SPAG5 的相互作用，流式细胞术显示ORP8 过表达后与空载体相比，人肝癌细胞系HepG2
的S 期细胞数增加，G1 期细胞数减少。结论：SPAG5 是与ORP8 相互作用的蛋白质，ORP8 过表达影响人肝癌细胞系HepG2 的细
胞周期，可能通过SPAG5 起作用。ORP8-SPAG5 的相互作用为进一步研究ORP8 在肝细胞中的功能及其机制提供了有用的线索 |
| 英文摘要: |
| Objective: To investigate the function of oxyterol binding protein related protein 8（ORP8）and to provide clues for better
understanding its function and mechanism. Methods: Yeast two-hybrid system screening human universal cDNA library was used to
obtained proteins interacting with ORP8. Putative interactions were further confirmed by GST Pull-down and Co-IP experiments. The
effects of ORP8 on cell cycle were detected by flow cytometry in HepG2 cells. Results: ORP8 interacted with SPAG5 in two-hybrid
screen using ORP8 as bait. ORP8-SAPG5 interaction was further confirmed in GST Pull-down and Co-IP experiments. Transient
over-expression of ORP8 in HepG2 cells decreased cell population of G0/G1-phase and increased S-phase compared with that in control.
Conclusion: ORP8 physically interact with SPAG5 and influence the cell cycle of HepG2 cell lines, probably through a mechanism involving
SPAG5. ORP8-SPAG5 interaction provides clues to further explore the function of ORP8 and its mechanism. |
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