Petitjean Audrey, Cavard Catherine, Shi Hong, Tribollet Violaine, Hainaut Pierre, Caron de Fromentel Claude
International Agency for Research on Cancer, 150 cours A Thomas, 69372 Lyon Cedex 08, France.
Oncogene. 2005 Jan 13;24(3):512-9. doi: 10.1038/sj.onc.1208215.
The TP63 gene, a member of the TP53 gene family, encodes several isoforms with (TAp63) or without (DeltaNp63) transactivating properties. Whereas the role of p63 in the normal development of squamous epithelia is well established, its function in other cell types remains to be elucidated. Here, we have analysed the expression of TA and DeltaNp63 isoforms in liver cells, by using both primary hepatocytes from wild type and p53-null mice and three human hepatocellular carcinoma (HCC) cell lines, according to the transformation state and the TP53 status of the cells. We observed the expression of DeltaNp63 isoforms only in a p53-null context. On the other hand, the expression of TAp63 isoforms was restricted to the HCC cell lines, whatever the TP53 status. We then studied the expression of TP63 upon genotoxic treatment. When treated with UVB or H(2)O(2), hepatocytes did not exhibit any change in p63 mRNA level. At the opposite, upon treatment with topoisomerase II inhibitors (doxorubicin or etoposide), the expression of TAp63 isoforms was clearly induced, independently of the TP53 status of cells. The same treatment did not induce any variation in the expression of DeltaNp63 isoforms, both at mRNA and protein levels. In HCC cell lines, doxorubicin or etoposide treatment also resulted in an increase of TAp63 transcripts only. This increase was accompanied by an increase in the intracellular level of TAp63 alpha protein. In parallel, we observed an upregulation of some p53-target genes related to cell cycle regulation, such as WAF1/CIP1, PIG3, 14-3-3sigma or GADD45, independently of the TP53 status of cells. In conclusion, we report for the first time that TA and DeltaNp63 alpha proteins are present in liver cells. Furthermore, our results suggest that p63 may partially substitute for wild-type p53, in counteracting uncontrolled liver cell proliferation in response to certain forms of DNA-damage.
TP63基因是TP53基因家族的成员之一,编码具有(TAp63)或不具有(DeltaNp63)反式激活特性的几种亚型。虽然p63在鳞状上皮正常发育中的作用已得到充分证实,但其在其他细胞类型中的功能仍有待阐明。在此,我们根据细胞的转化状态和TP53状态,使用野生型和p53基因敲除小鼠的原代肝细胞以及三种人肝细胞癌(HCC)细胞系,分析了TA和DeltaNp63亚型在肝细胞中的表达。我们仅在p53基因敲除的情况下观察到DeltaNp63亚型的表达。另一方面,无论TP53状态如何,TAp63亚型的表达都局限于HCC细胞系。然后,我们研究了基因毒性处理后TP63的表达。用UVB或H₂O₂处理时,肝细胞的p63 mRNA水平没有任何变化。相反,在用拓扑异构酶II抑制剂(阿霉素或依托泊苷)处理后,TAp63亚型的表达明显被诱导,与细胞的TP53状态无关。相同的处理在mRNA和蛋白质水平上均未诱导DeltaNp63亚型表达的任何变化。在HCC细胞系中,阿霉素或依托泊苷处理也仅导致TAp63转录物增加。这种增加伴随着TAp63α蛋白细胞内水平的增加。同时,我们观察到一些与细胞周期调控相关的p53靶基因上调,如WAF1/CIP1、PIG3、14-3-3σ或GADD45,与细胞的TP53状态无关。总之,我们首次报道TA和DeltaNp63α蛋白存在于肝细胞中。此外,我们的结果表明,p63可能部分替代野生型p53,以应对某些形式的DNA损伤,对抗不受控制的肝细胞增殖。