DeltaNp73alpha inhibits PTEN expression in thyroid cancer cells.
DeltaNp73 is a N-terminally truncated p53 family member with a dominant negative function, which is upregulated in cancer. PTEN is a lipid phosphatase, which is involved in the attenuation of tyrosine kinase signaling. PTEN expression is increased by p53, and its function is blunted in several malignancies. Because in most of the thyroid carcinomas, DeltaNp73alpha is upregulated, whereas PTEN expression down regulated, we investigated whether DeltaNp73alpha may influence PTEN expression in this cell model. We found that DeltaNp73alpha overexpression in thyroid cancer cells reduces PTEN expression, whereas DeltaNp73alpha down-regulation by siRNA increases PTEN expression. Real-time PCR indicated that overexpression of DeltaNp73alpha is able to reduce PTEN mRNA levels. Moreover, chromatin immunoprecipitation (ChIP) and luciferase assays indicated that DeltaNp73alpha binds to -1031-779 region of the PTEN promoter, which is a different site than that for p53, thereby inhibiting promoter activity. Interestingly, also the transcriptionally active p73 isoforms (TAp73alpha and TAp73beta) bound to this DNA sequence and, at variance with DeltaNp73alpha, stimulated PTEN promoter activity to an extent similar to that of p53. In accordance with its effect on PTEN protein levels, DeltaNp73alpha increased phospho-Akt protein content and, as a consequence, Mdm2-mediated p53 degradation. This effect of DeltaNp73alpha resulted in increased thyroid cancer cell proliferation and reduced apoptosis and was reverted by the PI3-kinase inhibitor LY294002, indicating the role of Akt pathway in this effect. Taken together, these results indicate a novel p73 regulated mechanism for PTEN expression in thyroid cancer cells, and that, also through this mechanism, DeltaNp73alpha exerts its protumorigenic effect.
Vella
Veronica
V
Department of Internal Medicine, Endocrinology Unit, University of Catania, Catania, Italy.
Puppin
Cinzia
C
Damante
Giuseppe
G
Vigneri
Riccardo
R
Sanfilippo
Mariangela
M
Vigneri
Paolo
P
Tell
Gianluca
G
Frasca
Francesco
F
D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival; D004268:DNA-Binding Proteins / Q000502:physiology*; D015536:Down-Regulation; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000502:physiology*; D051059:PTEN Phosphohydrolase / Q000037:antagonists & inhibitors / Q000235:genetics*; D011401:Promoter Regions, Genetic; D013964:Thyroid Neoplasms / Q000235:genetics* / Q000473:pathology; D016159:Tumor Suppressor Protein p53 / Q000032:analysis; D025521:Tumor Suppressor Proteins / Q000502:physiology*
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human