Figure 3

Caspase8 expression and NF-κB expression are correlated in patient samples. (a) Caspase8 mRNA expression quantified in subtypes of ovarian cancer as defined by the AOCS (average of probe sets 207686_at and 213373_s_at from Geo Dataset GSE9899), is highest in the C2 and C4 subtypes (P=7.5×10⁻⁶, based on t-test between groups C2+C4 versus others). (b) Patient sample subgroups were ranked by average expression of NF-κB signature. Caspase8 relative expression is shown for each patient sample, demonstrating highest expression in C2 and C4 subgroups, and lowest in C5. (c) Caspase8 expression correlates with NF-κB gene signature expression in AOCS patient samples from subtypes C2 and C5 (P=0.0002). (d) High versus Low expression of either Caspase8 or NF-κB gene signature was defined by median value across the entire data set. Shown are the proportions of samples within each subgroup with High (above the median) or Low (below the median) expression of either Caspase8 or NF-κB gene signature. (e) Overall survival of patients in each Caspase8/NF-κB category as defined in d. Patients with elevated expression of both Caspase8 and NF-κB had the longest median overall survival (P=0.03). (f) Progression-free survival of patients in each Caspase8/NF-κB category as defined in d. (g) Immunohistochemistry (IHC) average intensity scores of tumor cells expressing Caspase8, NF-κB-p65 and CD3 protein levels in AOCS tissue microarrays containing representative subtypes (C1, n=17; C2, n=19; C3, n=9; C4, n=20; C5, n=19). Percent positive cells were scored as 0 (<10%), 1 (10–24%), 2 (25–50%), 3 (50–74%) or 4 (75–100%). Staining intensity was scored as 0 (neg), 1 (weak), 2 (moderate), 3 (positive). In addition, tumors were scored for nuclear and cytoplasmic stain according to the above rubric. A composite score was tabulated by multiplying intensity score by percent positive score, and reported as average for each of the AOCS subtypes in the tissue microarrays. (h) Representative images of patient tissue from C2 subtype, showing high CASPASE 8, nuclear NF-κB-p65 and T-cell infiltrate, contrasted with C5 subtype showing comparatively low markers.