转录因子p53与AP-2基因家族成员AP-2beta发生突变后, 均会导致个体出现相应的疾病。本文首先利用两个重组质粒pCMV-HA-p53和pMyc-AP-2beta, 转染永生化的胚胎肾细胞HEK293(野生型), 在细胞中表达相应蛋白质。通过免疫共沉淀实验证明AP-2beta和p53蛋白在体内可以相互作用。并将梯度增加的pMyc-AP-2beta质粒及等量pCMV-HA-p53质粒转染到HEK293细胞, SDS-聚丙烯酰胺凝胶电泳实验证明AP-2beta能正向调控p53蛋白的表达。为了进一步探讨AP-2beta与p53的作用机制, 利用蛋白质合成抑制剂CHX(cycloheximide)处理转染了AP-2beta与p53表达质粒的细胞, 实验结果说明, AP-2beta能增加p53蛋白的稳定性来调控p53的表达。

The mutation of transcription factor p53 and AP-2beta which is AP-2 gene family member will result in individual diseases. Firstly, the use of two recombinant plasmids pCMV-HA-p53 and pMyc-AP-2beta to transfect immortalized embryonic kidney cells HEK293 (wild-type). Co-immunoprecipitation experiments showed that AP-2beta and p53 proteins may interact in vivo, And then the gradient of increasing pMyc-AP-2beta and the same amount of plasmid pCMV-HA-p53 were transfected into HEK293 cells.The results showed that AP-2beta can regulates of p53 positively by Western-blot.To further investigate the mechanism of interation of AP-2beta and p53, cells which were co-transfected AP-2beta and p53 plasmid were incubated with cycloheximide (CHX), experiments showed that AP-2beta can increased the stability of p53 proteins.