CNR1激活FIS1促进线粒体裂解抑制骨肉瘤细胞恶性进展  

Activation of FIS1 by CNR1 Promotes Mitochondrial Cleavage to Inhibit Malignant Progression of Osteosarcoma

作　　者：王良铭[1] 张小路[1] WANG Liangming;ZHANG Xiaolu(Department of Orthopedics,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院骨科,泉州362000

出　　处：《中国细胞生物学学报》2024年第12期2037-2044,共8页Chinese Journal of Cell Biology

基　　金：福建医科大学启航基金(批准号:2020QH1114)资助的课题。

摘　　要：该文探讨了大麻素受体1(cannabinoid receptor-1,CNR1)在骨肉瘤细胞恶性进展中的作用机制。通过生信分析CNR1在肿瘤组织中表达水平高低和远期生存率变化;采用CNR1过表达慢病毒感染骨肉瘤细胞;通过CCK-8、划痕实验、克隆实验、Transwell细胞侵袭实验和流式细胞凋亡实验检测骨肉瘤细胞增殖、集落形成和侵袭能力变化以及细胞凋亡水平;使用透射电镜、MitoSOX探针和免疫印迹检测线粒体形态和功能变化;通过裸鼠荷瘤实验验证CNR1高表达骨肉瘤细胞在体内的生长情况。结果显示CNR1在骨肉瘤组织中表达上调,并且CNR1高表达组患者的远期生存率显著高于低表达组;免疫印迹证实了慢病毒感染成功地诱导了骨肉瘤细胞系CNR1高表达;CNR1高表达显著抑制了骨肉瘤细胞增殖、集落形成和侵袭,并诱导了骨肉瘤细胞凋亡;CNR1激活了线粒体裂变蛋白1(mitochondrial fission 1 protein,FIS1)从而引起了线粒体过度裂解,诱发了线粒体途径凋亡;在载瘤裸鼠体内,CNR1过表达通过促进线粒体途径凋亡从而抑制骨肉瘤细胞恶性进展。The article explored the mechanism of CNR1 on the malignant progression of osteosarcoma cells.The changes of high and low CNR1 expression and long-term survival in tumor tissues were analyzed by bioanalysis;CNR1 overexpression lentivirus was used to infect osteosarcoma cells;CCK-8,scratch assay,clonogenic assay,Transwell cell invasion assay and flow apoptosis assay were used to detect the changes of osteosarcoma cells’proliferation,colony formation and invasion ability,as well as the level of apoptosis;transmission electron microscopy was used,MitoSOX probe and immunoblotting to detect changes in mitochondrial morphology and function;and nude mice loaded tumor assay to verify the growth of CNR1 high-expression osteosarcoma cells in vivo.The results showed that CNR1 expression was up-regulated in osteosarcoma tissues,and the long-term survival rate of patients in the group with high CNR1 expression was significantly higher than that in the group with low CNR1 expression;the immunoblotting confirmed that lentivirus infection successfully induced the high CNR1 expression in osteosarcoma cell lines;the high expression of CNR1 significantly inhibited osteosarcoma cells’proliferation,colony formation,and invasive ability and induced osteosarcoma cells’apoptosis;CNR1 activated FIS1 thereby causing excessive mitochondrial cleavage and inducing mitochondrial pathway apoptosis;in tumor-bearing nude mice,CNR1 overexpression inhibited malignant progression of osteosarcoma cells by promoting mitochondrial pathway apoptosis.

关 键 词：线粒体分裂 CNR1 凋亡 骨肉瘤 远期生存率 

分 类 号：R738.1[医药卫生—肿瘤]