目的 探讨SIRT6对肿瘤重要调节分子胰岛素样生长因子2(IGF2)的转录后调控机制.方法 在工具细胞系293T细胞中过表达SIRT6以及应用免疫共沉淀(IP)方法富集所有可能与SIRT6相互作用的蛋白,行质谱实验鉴定,并进一步通过IP/Western blot方法,检测SIRT6和IGF2 mRNA结合蛋白(IMP1)的相互作用,以及 SIRT6对IMP1的乙酰化/磷酸化修饰水平的影响.结果 质谱结果证明SIRT6和IMP1存在相互作用,并通过构建SIRT6和IMP1的过表达载体,进一步验证了SIRT6与IMP1的直接相互作用.检测乙酰化和磷酸化水平发现,SIRT6并未直接影响IMP1的乙酰化水平,而是促进其磷酸化修饰.结论 SIRT6可能通过促进对IMP1的磷酸化而起到在转录后水平抑制细胞中IGF2 mRNA的作用.%Objective To investigate the post-transcriptional regulation mechanism of insulin-like growth factor 2 (IGF2),which is an important tumor regulator,by SIRT6. Methods SIRT6 was overexpressed in 293T cells and IP was used to enrich SIRT6 and mass spectrometry(MS)was used to detect the protein that interacted with SIRT6. Western blot was used to validate the interacted protein and its acetylation/phosphorylation modification status. Results SIRT6 did not change the acetylation modification status of IMP1,but the phosphorylation status of IMP1 was elevated in the presence of SIRT6. Conclusions SIRT6 may regulate IGF2 though promoting the phosphoryla-tion of IMP1 in 293T cells.
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