不同状态p53小鼠骨髓间质干细胞分泌外泌体的差异

Effects of different states of p53 on the exosomes secreted by mouse bone marrow mesenchymal stem cells

目的:比较3种p53状态(p53^(+/+)、p53^(+/-)和p53^(-/-))的小鼠骨髓间质干细胞(mouse bone marrow mesenchymal stem cells,m BMMSC)分泌的外泌体(exosome)是否存在差异。方法:采用透射电镜及蛋白质印迹鉴定和分析p53不同状态m BMMSC分泌的外泌体的形态及膜表面特异性标志蛋白CD9和CD63,采用Nanosight纳米颗粒跟踪分析仪比较外泌体浓度和大小的差异。结果:3种m BMMSC分泌的外泌体均为圆形或椭圆形的膜性囊泡,直径均在100 nm左右。p53^(-/-)m BMMSC和p53^(+/-)m BMMSC分泌的外泌体浓度明显高于p53^(+/+)m BMMSC,p53^(-/-)m BMMSC与p53^(+/-)m BMMSC分泌的外泌体浓度无显著性差异。3种细胞来源的外泌体均表达膜表面特异性标志蛋白CD9、CD63。结论:p53缺失的m BMMSC能分泌更多的外泌体。

Objective： To detect the difference of exosomes derived from p53 ＋/＋ mBMMSC, p53 ＋/- mBMMSC and p53 -/- mBMMSC. Methods ： The purified exosomes derived from p53 ＋/＋ mBMMSC,p53 ＋/- mBMMSC and p53-/- mBMMSC were identified by transmission electron microscopy for morphology and western blotting assay revealed the expression levels of exosomal markers CD9 and CD63. Nanoparticle Tracking Analysis （NTA） was used to detected the difference of concentration and diameter in exosomes de- rived from p53 ＋/＋ mBMMSC, p53 ＋/- mBMMSC and p53 -/- mBMMSC. Results： Exosomes derived from p53 ＋/＋ mBMMSC ,p53 ＋/-mBMMSC and p53-/-mBMMSC were all lipid bilayer membrane vesicles whose sizes were about 100 nm. The concentration of exosomes derived from p53 ＋/-mBMMSC and p53-/- mBMMSC were significantly more than p53 ＋/＋ mBMMSC while p53 ＋/-mBMMSC and p53-/-mBMMSC had no significant difference. In addition, the diameter of exosomes derived from p53 ＋/＋ mBMMSC, p53 ＋/- mBMMSC and p53-/- mBMMSC was similar. All exosomes were expressed exosomal markers CD9 and CD63. Conclusion ： Mouse bone marrow mesenchymal stem cells lost p53 gene cloud release more exosomes.