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. 2010 Jul 1;120(8):2842–2857. doi: 10.1172/JCI36125

Figure 2. DAB2 promoter methylation correlates with low-level expression and predicts metastasis and poor clinical outcome in SCC.

Figure 2

(A) Analysis of DAB2 promoter methylation by MSP analysis in 9 HNSCC primary tumor isolates. (B) DAB2 promoter methylation correlates with low-level DAB2 expression. Analysis of DAB2 expression by RT-PCR in 2 normal (N) and 5 primary HNSCC tumor (T) samples. GAPDH RT-PCR is shown as a loading control. (C) DAB2 promoter methylation correlates with metastasis in VSCC. Graphical representation of DAB2 MSP analysis of 26 matched normal tissue and primary tumors isolated from VSCC patients. Primary tumor data is separated into patients with (16 patients) and without (10 patients) inguinal nodal involvement, and analysis of matched metastasis material is also shown. (D and E) DAB2 promoter methylation predicts poor clinical outcome in HNSCC. Log-rank survival analysis in HNSCC patients. The Kaplan-Meier curves show analyses of overall survival (D) and progression-free survival (E) in patients with HNSCC, as a function of DAB2 promoter methylation (methylated) or lacking methylation (unmethylated). The numbers under the graphs indicate the numbers of cases analyzed. (D) Overall survival is significantly worse in patients with tumors with methylation in the DAB2 promoter (P < 0.0001). (E) Progression-free survival is significantly worse in patients with tumors with methylation in the DAB2 promoter (P < 0.0001). (F) MSP analysis correlates with pyrosequencing analysis. HNSCC tumor samples were analyzed by MSP analysis as in A and by quantitative pyrosequencing of CpG dinucleotides 39–44 (depicted in Figure 1D). Data represent the average percentage of methylation at each CpG, and box plots are shown for MSP+ve and MSP–ve samples. (G) MSP analysis correlates with DAB2 expression levels. qRT-PCR analysis of DAB2 expression of samples is analyzed as in F. Expression levels are represented in relation to the highest expressing sample, which is assigned the arbitrary value of 1. (F and G) Data are analyzed by nonparametric Mann-Whitney statistics. In box plots, the 75th and 25th percentiles are represented by the top and bottom of the box, respectively. The horizontal lines refer to the mean. **P < 0.01.