【摘 要】
：
E3泛素化连接酶TRIM21(tripartite motif containing 21)在不同类型肿瘤中扮演癌基因或抑癌基因的角色,从而影响细胞功能及临床预后.然而,TRIM21在下咽癌中的生物学功能和分子机制尚不清楚.本文采用RT-PCR和Western印迹技术检测了TRIM21在下咽癌组织中的表达水平,采用基因过表达和RNA干扰技术分析了TRIM21过表达和基因沉默对细胞增殖、迁移和分化的影响,利用蛋白质免疫共沉淀联合质谱鉴定TRIM21互作蛋白质,并进一步研究TRIM21影响下咽癌分化的分子机制
【机 构】
：
山西医科大学基础医学院生物化学与分子生物学教研室,太原030001;山西医科大学细胞生理学教育部重点实验室,太原030001;青岛大学附属医院耳鼻咽喉头颈外科 青岛266003;青岛大学附属医院耳鼻咽
论文部分内容阅读
E3泛素化连接酶TRIM21(tripartite motif containing 21)在不同类型肿瘤中扮演癌基因或抑癌基因的角色,从而影响细胞功能及临床预后.然而,TRIM21在下咽癌中的生物学功能和分子机制尚不清楚.本文采用RT-PCR和Western印迹技术检测了TRIM21在下咽癌组织中的表达水平,采用基因过表达和RNA干扰技术分析了TRIM21过表达和基因沉默对细胞增殖、迁移和分化的影响,利用蛋白质免疫共沉淀联合质谱鉴定TRIM21互作蛋白质,并进一步研究TRIM21影响下咽癌分化的分子机制.结果 表明,TRIM21在中高分化下咽癌中高表达,提示TRIM21可能调控肿瘤细胞分化.在FaDu细胞中,过表达或shRNA干扰TRIM21可分别抑制和促进细胞增殖和迁移.此外,过表达TRIM21后,分化标志分子角蛋白10 (keratin 10,KRT10)、内披蛋白(involucrin,IVL)和谷氨酰胺转氨酶1(transglutaminase 1,TGM1)蛋白质表达水平增高,而shRNA干扰TRIM21后,上述蛋白质表达下降.质谱鉴定及蛋白质生物信息分析提示,TRIM21可能与细胞骨架蛋白质调控密切相关.进一步验证表明,TRIM21与角蛋白10相互作用.蛋白质合成抑制剂环已酰亚胺处理细胞后,与对照相比,TRIM21过表达细胞中角蛋白10表达增高,且过表达TRIM21可升高角蛋白10的泛素化水平.综上所述,我们的研究表明,TRIM21可能通过介导细胞骨架蛋白质泛素化修饰,提高蛋白质稳定性促进下咽癌分化.
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