BACKGROUND.Although p53 overexpression is frequent in head and neck squamous cell carcinomas (HNSCCs), controversy remains regarding the prognostic significance of that overexpression. The objective of this study was to investigate the expression pattern and prognostic significance of p53 expression in HNSCC of the same location, treated in the same way, and with long‐term follow‐up.METHODS.P53 expression was determined by immunohistochemistry in paraffin‐embedded tissue specimens from 107 consecutive patients (107 primary squamous cell carcinomas of the supraglottic larynx and 46 matched lymph node metastases). All patients underwent surgical resection and bilateral neck dissection.RESULTS.A strong correlation was observed between p53 expression in the primary tumor and in the matched lymph node metastases (P = .0001). P53 overexpression in the lymph nodes was an independent predictor of regional recurrence (P = .027). Likewise, expression of p53 in the lymph nodes correlated significantly with disease‐specific survival (P = .018). Five years after treatment, 70% of patients with p53‐negative, metastatic lymph nodes remained alive, whereas only 30% of patients with p53‐positive lymph nodes remained alive. In multivariate analysis, lymph node status and p53 expression in the lymph nodes remained associated with survival.CONCLUSIONS.The current data suggested that, although p53 overexpression is common in supraglottic carcinomas, its expression in the primary tumor is of limited clinical significance. However, the results supported the role of p53 in the lymph node metastases as an independent predictor of regional failure and a poor prognosis in patients with HNSCC. A prospective trial is indicated to validate these findings. Cancer 2007. © 2007 American Cancer Society.