Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5642553 | Oral Oncology | 2017 | 11 Pages |
â¢ÎNp63β negatively regulates Wnt5a, Ror2 and MMP-2 expression in tongue SCC cell lines.â¢Wnt5a-Ror2 signaling affects tongue SCC cell motility and production of MMP-2.â¢Wnt5a or Ror2 expression is strongly involved in lymph node metastasis of tongue SCC.â¢Tongue SCC patients with high Wnt5a or Ror2 expression show poorer prognosis.â¢Wnt5a-Ror2 signaling enhances tongue SCC cell aggressiveness via ÎNp63β regulation.
ObjectivesWe previously showed that ÎNp63β, a splicing variant of ÎNp63, mediated EMT and affected cell motility. DNA microarray was thus performed to elucidate the mechanism that ÎNp63β affects cell motility. As the results, Wnt5a was significantly down-regulated by ÎNp63β overexpression in tongue SCC cell line (SQUU-B) with EMT phenotype.Materials and methodsSeven OSCC cell lines were used. Expression of ÎNp63, Wnt5a, its receptor Ror2, and matrix metalloproteinases (MMPs) were analyzed by RT-PCR, real-time PCR, and western blotting, and gelatin zymography. Furthermore, we examined the effects of siRNA for Wnt5a or Ror2 and recombinant human Wnt5a (rhWnt5a) on motility of tongue SCC cells. Biopsy specimens from tongue SCC patients were used for immunohistochemical staining of Wnt5a and Ror2.ResultsWnt5a and Ror2 were expressed only in SQUU-B cells without ÎNp63 expression, and negatively associated with ÎNp63 expression in other cells. ÎNp63β overexpression in SQUU-B cells decreased Wnt5a and Ror2 expression. By Wnt5a or Ror2 knockdown, cell motility was remarkably inhibited, but EMT markers expression was unaffected. MMP-2 expression and the activities inversely correlated with ÎNp63 expression, and were inhibited by Wnt5a or Ror2 knockdown. Cell motility and MMP-2 activities were recovered by adding rhWnt5a in the cells with Wnt5a knockdown, but not in those with Ror2 knockdown. Moreover, immunohistochemical analyses in tongue SCC specimens found that high expression of Wnt5a or Ror2 was associated with poorer prognosis.ConclusionWnt5a-Ror2 signaling enhanced tongue SCC cell aggressiveness and promoted production of MMP-2 following ÎNp63β-mediated EMT.