Standard-protocol IHC was done on Human Lung Cancer Survey tissue microarrays (US Biomax, Milan, Italy) and formalin-fixed paraffin-embedded (FFPE) XT tissues. The UltraTek HRP Anti-Polyvalent (DAB) Staining System (ScyTek, Logan, UT, USA) was used for blocking and detection; apoptosis was detected with TUNEL-POD kit (Roche, Monza, Italy). Labeling was detected with the UltraTek HRP Anti-Polyvalent (DAB) Staining System (ScyTek). Stained sections were examined with a Leica DM1000 microscope (Leica Microsystems Srl, Milan, Italy), photographed with a Jenoptik ProgRes SpeedXT Core 3 camera (Jenoptik AG, Vienna, Austria), and analyzed with ProgRes Capture Pro 2.8 software (Jenoptik AG).
Protein expression was evaluated with histoscore method (H-score): staining intensity was rated from 0 (negative) to 3 (strong) for each sample by two independent observers and the mean was used. H-score was calculated multiplying the number of cells within each category in at least three different × 200 fields. Human protein KI-67 expression and TUNEL scores were reported as percentages of labeled cells with respect to total cells.
Protein expression was evaluated with histoscore method (H-score): staining intensity was rated from 0 (negative) to 3 (strong) for each sample by two independent observers and the mean was used. H-score was calculated multiplying the number of cells within each category in at least three different × 200 fields. Human protein KI-67 expression and TUNEL scores were reported as percentages of labeled cells with respect to total cells.