Article Figures & Data
Figures
- Figure 1.
Expression of total HER2, total HER3, HER2:HER3 heterodimers, and HER3–PI3K complexes is significantly higher in HPV+ compared with HPV− HNSCC tumors. Expression of total HER1 is significantly lower in HPV− HNSCC. Box-and-whisker plots of VeraTag variables by p16 (HPV) status. P values are for a Wilcoxon test comparing p16 positive with p16 negative.
- Figure 2.
Expression of EGFR/HER1, HER2, and HER3 in the HNSCC TCGA cohort. Levels of HER2 and HER3 mRNA are significantly higher in HPV+ tumors and expression of EGFR/HER1 is significantly higher in HPV− HNSCC tumors as assessed by RNAseq (HPV positive n = 36; HPV negative n = 240. Box-and-whisker plots of EGFR/HER1, HER2, and HER3 mRNA by HPV status from TCGA. Expression levels have been converted to Z scores, P values are for a Wilcoxon test comparing HPV+ and HPV−.
- Figure 3.
EGFR/HER1 VeraTag expression levels positively correlate with IHC results. A, HER1 dimer plotted against HER1 total (HER1/EGFR by VeraTag) and B, total EGFR (HER1/EGFR by conventional IHC). HER1 dimer was positively correlated with both HER1 total by VeraTag and EGFR IHC. Curves and statistics are from a least-squares regression on a 3-knot restricted cubic spline fit for the variables on the x-axis. R2 reflects the total variation explained by the regression model.
- Figure 4.
Afatanib is significantly more effective at inhibiting cell growth in cetuximab-resistant and HPV+ HNSCC cell lines. A, Cal33 vector transfected control cells were treated with C225 (cetuximab) or afatinib for 72 hours followed by cell viability assessment. There was no significant difference in the effects of C225 or afatinib in these cells. Cal33vIII (B), UMSCC-47 (C), or UPCI-90 (D) was treated with C225 or afatinib for 72 hours, with afatinib demonstrating significant effects on inhibiting cell viability compared with C225.
- Figure 5.
Afatinib abrogates EGFR/HER2 signaling in HNSCC models. HNSCC cell lines, including A, HPV− cetuximab-sensitive (Cal33 VC), B, HPV− cetuximab-resistant (Cal33 vIII), and C, HPV+ (UMSCC-47, UPCI SCC-90) HNSCC models were treated with increasing concentrations of afatinib. After 24 hours of treatment, cells were harvested to obtain cell lysates. Forty micrograms of protein/lane was subjected to gel electrophoresis and probed with pHER2Tyr1248, pAkt, total HER2, and total Akt. β-Tubulin was used as a loading control. Phosphorylated protein was compared with total protein, and the ratio is listed under each protein band. The experiment was repeated three times with similar results.
Tables
- Table 1.
HNSCC patient clinical and pathologic characteristics
Patient characteristic N = 88 (%) Age at diagnosisa Median 59 Range 23–90 Gender Male 68 (79) Female 28 (21) Clinical T stage 1 13 (15) 2 24 (27) 3 10 (11) 4 15 (17) Recurrent/unknown 26 (30) Clinical N stage 0 17 (19) 1 12 (14) 2 32 (36) 3 3 (3) Recurrent/unknown 24 (27) Siteb Oral cavity 24 (28) Oropharynx 30 (35) Hypopharynx 4 (5) Larynx 16 (19) Nasopharynx 3 (3) Other (parotid, skin, neck) 3 (3) Histologyc Squamous cell 71 (83) Basaloid squamous cell 9 (10) Other 6 (7) P16d Positive 34 (41) Negative 48 (59) Cancer typee Primary 66 (78) Recurrence 19 (22)
Supplementary Data
- Supplementary Methods - Detailed methods for VeraTag assays.
Volume 21, Issue 20
