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目的探讨组蛋白甲基转移酶SETD2对P53功能的调控。方法 PCR扩增人源SETD2并重组入真核表达载体;通过间接免疫荧光鉴定SETD2的亚细胞定位;通过GST Pull-down实验寻找P53结合SETD2的区域;流式细胞术分析SETD2对P53参与细胞凋亡的影响。结果成功克隆了人源SETD2的全长,并将其成功构建到真核表达载体中;SETD2以胞核定位为主;体外直接相互作用实验证实SETD2和P53存在相互作用,SETD2分子中部的SET结构域和C端都可以结合P53;流式细胞分析结果表明SETD2对于P53参与的细胞凋亡具有明显的促进作用。结论本研究揭示P53正向调控因子SETD2对P53介导的细胞凋亡具有促进效应,从而将组蛋白表遗传学修饰与P53转录因子活性调控关联起来,研究结果对肿瘤、免疫等疾病的防治具有一定的科学意义。
Objective To investigate the regulation of P53 function by histone methyltransferase SETD2. METHODS: Human SETD2 was amplified by PCR and inserted into eukaryotic expression vector. The subcellular localization of SETD2 was identified by indirect immunofluorescence. The region of P53 binding to SETD2 was found by GST pull-down assay. Flow cytometry The impact of death. Results The full length of human SETD2 gene was successfully cloned and constructed into eukaryotic expression vector. SETD2 was mainly located in nucleus. Direct interaction in vitro confirmed the interaction between SETD2 and P53. The SET structure in the middle of SETD2 Both P53 and C53 could bind to P53. Flow cytometry analysis showed that SETD2 could significantly promote the apoptosis of P53 cells. Conclusions This study reveals that SETD2, a positive regulator of P53, has a promoting effect on P53-mediated apoptosis and thus correlates histone modifications of histones with the regulation of P53 transcription factor activity. The results of this study have shown that the prevention and treatment of tumors, Certain scientific significance.