Institute of Medical Biology, A*STAR, Singapore.
PLoS Pathog. 2011 Sep;7(9):e1002256. doi: 10.1371/journal.ppat.1002256. Epub 2011 Sep 29.
Cervical carcinomas result from cellular transformation by the human papillomavirus (HPV) E6 and E7 oncogenes which are constitutively expressed in cancer cells. The E6 oncogene degrades p53 thereby modulating a large set of p53 target genes as shown previously in the cervical carcinoma cell line HeLa. Here we show that the TAp63β isoform of the p63 transcription factor is also a target of E6. The p63 gene plays an essential role in skin homeostasis and is expressed as at least six isoforms. One of these isoforms, ΔNp63α, has been found overexpressed in squamous cell carcinomas and is shown here to be constitutively expressed in Caski cells associated with HPV16. We therefore explored the role of p63 in these cells by performing microarray analyses after repression of endogenous E6/E7 expression. Upon repression of the oncogenes, a large set of p53 target genes was found activated together with many p63 target genes related to cell adhesion. However, through siRNA silencing and ectopic expression of various p63 isoforms we demonstrated that TAp63β is involved in activation of this cell adhesion pathway instead of the constitutively expressed ΔNp63α and β. Furthermore, we showed in cotransfection experiments, combined with E6AP siRNA silencing, that E6 induces an accelerated degradation of TAp63β although not through the E6AP ubiquitin ligase used for degradation of p53. Repression of E6 transcription also induces stabilization of endogenous TAp63β in cervical carcinoma cells that lead to an increased concentration of focal adhesions at the cell surface. Consequently, TAp63β is the only p63 isoform suppressed by E6 in cervical carcinoma as demonstrated previously for p53. Down-modulation of focal adhesions through disruption of TAp63β therefore appears as a novel E6-dependent pathway in transformation. These findings identify a major physiological role for TAp63β in anchorage independent growth that might represent a new critical pathway in human carcinogenesis.
宫颈癌是由人类乳头瘤病毒(HPV)E6 和 E7 癌基因引起的细胞转化引起的,这些癌基因在癌细胞中持续表达。E6 癌基因降解 p53,从而调节大量的 p53 靶基因,如先前在宫颈癌细胞系 HeLa 中所示。在这里,我们表明 p63 转录因子的 TAp63β 同工型也是 E6 的靶标。p63 基因在皮肤稳态中发挥着至关重要的作用,并表达至少六种同工型。其中一种同工型ΔNp63α 在鳞状细胞癌中过度表达,并且在这里与 HPV16 相关的 Caski 细胞中持续表达。因此,我们通过抑制内源性 E6/E7 表达后进行微阵列分析来探索 p63 在这些细胞中的作用。在抑制癌基因后,发现一大组 p53 靶基因与许多与细胞黏附相关的 p63 靶基因一起被激活。然而,通过 siRNA 沉默和各种 p63 同工型的异位表达,我们证明 TAp63β 参与了该细胞黏附途径的激活,而不是持续表达的 ΔNp63α 和 β。此外,我们在共转染实验中结合 E6AP siRNA 沉默表明,E6 诱导 TAp63β 的快速降解,尽管不是通过用于降解 p53 的 E6AP 泛素连接酶。E6 转录的抑制也诱导宫颈癌细胞中内源性 TAp63β 的稳定化,导致细胞表面的焦点黏附物浓度增加。因此,如先前对 p53 所示,TAp63β 是 E6 在宫颈癌中抑制的唯一 p63 同工型。通过破坏 TAp63β 来下调焦点黏附似乎是转化中的一种新的 E6 依赖性途径。这些发现确定了 TAp63β 在无锚定生长中的主要生理作用,这可能代表人类致癌作用中的新关键途径。