Loss of PTEN during tumor transformation and lymph node involvement. (A) Representative examples of PTEN expression in normal, dysplastic, and HNSCC from humans demonstrating PTEN expression. Note PTEN down-regulation in the dysplastic epithelium and in the HNSCC epithelial cell invasion (black arrows). (B) Analysis of PTEN expression on oral epithelia (n = 10) and four TMAs for HNSCC (containing n = 709). In normal oral epithelia, PTEN staining was positive mostly in the basal layer. Note an overall PTEN loss in 31.2% of all HNSCC tumors (**P < .01). (C) Representative examples of PTEN staining in HNSCC TMA tumor core (n = 709; positive, top; negative, bottom). (D) Scatter plot of relative microarray signal intensities for PTEN mRNA in HNSCC samples from patients with negative lymph nodes (N0) versus presence of lymph node metastases (N1+; error bar, SEM; **P ≤ .01; ***P ≤ .001).