Gene expression signatures in HuH7 cells transfected with super hybrid p53 version 2.. Homo sapiens

The p53 tumor suppressor belongs to a gene family that includes two other structurally and functionally related members: p63 and p73. However, the regulation of p53 activity differs significantly from that of p73 and p63. To enhance the tumor suppressive activity of p53, we created a p63/p53 hybrid molecule, named p63-53O, which comprises the transcriptional activation and DNA-binding domains of TAp63γ and the oligomerization domain of p53. In this study, we generated a series of p63-53O derivatives and developed an advanced hybrid molecule named “Super Hybrid p53 (SHp53).” SHp53 efficiently transactivated several proapoptotic genes, including CASP10, KCNK3, and PYCARD, compared with p53. Moreover, the silencing of these proapoptotic genes partially abolished the apoptotic response to SHp53 in human cancer cells. The potency of SHp53 for suppressing tumorigenesis was also evaluated using in vivo models. Thus, our results identify a better p63/p53 hybrid molecule for the development of anti-cancer therapies. Overall design: HuH-7 human hepatocellular carcinoma cells were either transduced with adenoviral vectors expressing p53, TAp63gamma, p53-p63 hybrid (super hybrid p53 version 2, p63-53O-FL), or lacZ. After 24 hours, total RNA was isolated and analyzed by hybridization to Agilent-028004 SurePrint G3 Human GE 8x60K Microarray.

p53抑癌基因隶属于一个基因家族，该家族另有两个结构与功能均高度相关的同源成员：p63与p73。然而，p53的活性调控机制与p73、p63存在显著差异。为增强p53的抑癌活性，我们构建了一款p63/p53杂合分子，命名为p63-53O，其包含TAp63γ的转录激活结构域与DNA结合结构域，以及p53的寡聚化结构域。本研究中，我们构建了一系列p63-53O衍生物，并开发出一款高级杂合分子，命名为超级杂合p53（Super Hybrid p53，SHp53）。相较于野生型p53，SHp53可有效激活包括CASP10、KCNK3及PYCARD在内的多种促凋亡基因。进一步实验表明，沉默这些促凋亡基因可部分削弱人癌细胞中SHp53诱导的凋亡应答。我们还通过体内模型评估了SHp53抑制肿瘤发生的效能。综上，本研究成果鉴定出一款更优异的p63/p53杂合分子，可用于抗癌疗法的开发。

整体实验设计：将HuH-7人肝癌细胞分别用表达p53、TAp63γ、p53-p63杂合分子（超级杂合p53版本2，p63-53O-FL）以及lacZ的腺病毒载体进行转导。转导24小时后，提取总RNA，通过与Agilent-028004 SurePrint G3 Human GE 8x60K Microarray杂交进行分析。