E3泛素化连接酶TRIM29在骨肉瘤细胞生长中的调控机制研究OA北大核心CSTPCD
The regulation mechanism of E3 ubiquitin ligase TRIM29 in the growth of osteosarcoma cells
目的:E3泛素化连接酶TRIM29与肿瘤的恶性生长紧密相关,本研究旨在分析并探讨TRIM29在骨肉瘤中的表达及其潜在的分子机制.方法:通过蛋白质印迹法(Western blot)、免疫组化(IHC)及qRT-PCR分析骨肉瘤组织中TRIM29基因的表达情况,采用si-RNA质粒转染至骨肉瘤细胞U2-OS中,CCK8和EDU检测骨肉瘤细胞的增殖能力,进一步采用葡萄糖摄取量、乳酸生成量及G6P检测糖酵解水平变化并分析潜在机制,建立BACB/c裸小鼠皮下移植瘤模型,观察下调TRIM29观察对荷瘤瘤生长的影响.结果:qRT-PCR结果证实与正常组织相比,骨肉瘤组织中TRIM29的mRNA表达水平显著提升,进一步结果也证实其蛋白水平也同样显著提升.下调骨肉瘤细胞中TRIM29的表达后,CCK-8及EDU实验皆表明其增增殖能力减弱,相反其凋亡比例升高,细胞于G1区阻滞,其乳酸生成量、葡萄糖消耗、G6P也相应的减弱,糖酵解关节限速酶HKII、PFKFB3、PKM、LDHA及GLUT1等蛋白也相应的减低,同样体内实验也表明下调TRIM29的表达后,荷瘤鼠体积、重量也随之减轻,生长速度减慢.结论:TRIM29在骨肉瘤中高表达,抑制骨肉瘤细胞中TRIM29的表达后,其细胞增殖能力及糖酵解水平减低.降低TRIM29的表达通过抑制细胞糖酵解水平进而抑制骨肉瘤细胞的生长.
Objective:To analyze and explore the expression of TRIM29 in osteosarcoma and its potential molecular mecha-nism.E3 ubiquitination ligase TRIM29 is closely related to the malignant growth of tumors.Methods:The expression of TRIM29 gene in osteosarcoma was analyzed by Western blot,IHC and qRT-PCR,CCK8 and EDU were used to detect the value-added level of osteosarcoma cells after downregulation of TRIM29.Further,glucose uptake,lactate production and G6P were used to detect the changes of glycolysis level and analyze the underlying mechanism.A subcutaneous tumor model of BACB/C nude mice was established to observe the effect of downregulation of TRIM29 on the growth of transplanted tumors.Results:qRT-PCR re-sults confirmed that the mRNA expression level of TRIM29 in osteosarcoma tissues was significantly higher than that in normal tis-sues.Similarly,Western blot and IHC results also confirmed that the protein level of TRIM29 in osteosarcoma tissues was signifi-cantly higher.After downregulating the expression of TRIM29 in osteosarcoma cells,CCK-8 and EDU experiments both showed that its value-added ability was weakened,on the contrary,its apoptosis rate was increased.In the G1 phase of cells,its lactate pro-duction,glucose consumption,G6P were also correspondingly weakened,and the glycolytic joint rate limiting enzymes HKII,PFKFB3,PKM,LDHAA,GLUT1 and other proteins were also correspondingly reduced.Similarly,in vivo experiments also showed that after downregulating the expression of TRIM29,the weight and growth rate were reduced accordingly.Conclusion:TRIM29 is highly expressed in osteosarcoma cells.After inhibiting the expression of TRIM29 in osteosarcoma cells,its cell prolif-eration ability and glycolysis level are decreased.TRIM29 inhibits the growth of osteosarcoma cells by restraining the glycolysis level of cells
钟贞浩;文王强;李国军;史册;唐彦超;陈焕雄
海南医学院第一附属医院 海南省脊柱外科临床医学中心,海南 海口 570102徐州医科大学附属宿迁医院骨科,江苏徐州 223865北京大学第三医院骨科,北京 100083
临床医学
TRIM29糖酵解增殖骨肉瘤
TRIM29GlycolysisProlifessionOsteosarcoma
《海南医学院学报》 2024 (014)
1041-1047 / 7
This study was supported by Young Science and Technology Talents Innovation Program of Hainan Science and Technology Association(QCXM202014)海南省科协青年科技英才创新计划项目(QCXM202014)
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