RanBP9激活线粒体途径诱导骨肉瘤细胞凋亡  

Ran-binding protein 9 overexpression induces apoptosis in osteosarcoma cells by activating endogenous mitochondrial pathway

作　　者：郭栎枫 吕杨帆 曹亚 郭乔楠 尹良军[1] GUO Lifeng;LYU Yangfan;CAO Ya;GUO Qiaonan;YIN Liangjun(Department of Orthopedics,the Second Affiliated Hospital of Chongqing Medical University,Chongqing,400010;Department of Pathology,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037,China)

机构地区：[1]重庆医科大学附属第二医院骨科,重庆400010 [2]陆军军医大学(第三军医大学)第二附属医院病理科,重庆400037

出　　处：《第三军医大学学报》2020年第12期1174-1181,共8页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81672653,816902591)。

摘　　要：目的探讨过表达RanBP9对骨肉瘤细胞凋亡的作用及其机制。方法采用Real-time PCR、Western blot检测成骨细胞及不同骨肉瘤细胞株(SaOS2、MG63、U2OS和HOS)中RanBP9的mRNA和蛋白表达水平。使用慢病毒建立稳定过表达RanBP9的SaOS2骨肉瘤细胞模型,并利用流式细胞仪及TUNEL染色法检测过表达RanBP9组及对照组骨肉瘤细胞凋亡率,使用线粒体膜电位检测过表达RanBP9组及对照组细胞凋亡过程中发生的线粒体膜电位变化。结果RanBP9在成骨细胞和骨肉瘤细胞株中均呈阳性表达,在成骨细胞中的表达显著高于骨肉瘤细胞(P<0.05),并且在恶性程度较低的骨肉瘤细胞株MG63和HOS中的表达要高于恶性程度较高的细胞株SaOS2和U2OS(P<0.05)。过表达RanBP9降低骨肉瘤细胞的增殖能力,促进凋亡率上升(P<0.05)。过表达RanBP9明显增加Caspase-3和Cytc的表达水平,并同时上调Bax/Bcl-2的比率(P<0.05);过表达RanBP9破坏线粒体膜电位,诱导内源性线粒体凋亡信号转导途径。结论RanBP9通过上调内源性线粒体通路发挥诱导骨肉瘤细胞凋亡的作用。Objective To investigate the effect of overexpression of Ran-binding protein 9(RanBP9)on apoptosis of osteosarcoma cells and the underlying mechanism.Methods Real-time PCR and Western blotting were used to detect the mRNA and protein expression of RanBP9 in cultured osteoblasts and different osteosarcoma cell lines(SaOS2,MG63,U2OS and HOS).In a SaOS2 cell model with stable RanBP9 overexpression induced by transfection with a lentiviral vector,the cell apoptotic rate was determined using flow cytometry and TUNEL assay,and the changes in mitochondrial transmembrane potential were analyzed.Results RanBP9 was positively expressed in cultured osteoblasts and in the 4 osteosarcoma cell lines.RanBP9 expression was significantly higher in the osteoblasts than in the osteosarcoma cells,and was significantly higher in osteosarcoma cell lines MG63 and HOS than in SaOS2 and U2OS cell lines that had higher levels of malignancy.In SaOS2 cells,overexpression of RanBP9 obviously suppressed cell proliferation and promoted cell apoptosis and significantly increased the expression levels of caspase-3 and Cytc and the ratio of Bax/Bcl-2,resulting also in the dissipation of mitochondrial transmembrane potential and activation of the mitochondrial pathway to induce cell apoptosis.Conclusion RanBP9 promotes osteosarcoma cell apoptosis via the endogenous mitochondrial pathway.

关 键 词：骨肉瘤 RanBP9 凋亡 增殖 

分 类 号：R341[医药卫生—基础医学] R394.2