According to the IHC scoring system, 16 cases (8/20 NSCLC and 8/1

According to the IHC scoring system, 16 cases (8/20 NSCLC and 8/13 pulmonary mCRC) showed an intense EGFR-immunoCB-839 datasheet reactivity (score 3+) (fig 2), 5 moderate reactivity (score 2+) and 3 weak reactivity MEK inhibitor (score 1+). No immunoreactivity (score 0) was observed in 9 cases (7 NSCLC and 2 mCRC). In particular, among the 27 polysomic cases detected by CISH (12 low polysomy, 15 high polysomy), 17 (63%) scored 2+/3+ (6 NSCLC and

11 pulmonary mCRC), and 10 (37%) scored 0/1+ (8 NSCLC and 2 pulmonary mCRC). The 2 NCSLC amplified by CISH displayed a 3+ score. We did not observe any statistically significant correlation between IHC scores and CISH (p = .85). Figure 2 Immunocytochemical evaluation of EGFR on non small cell lung carcinoma. Immunohistochemistry for EGFR in large cell carcinoma (LCC) FNAC cell block evidencing a strong membrane immunoreactivity (score 3+). Original magnification ×400. Furthermore, a comparison between CISH and FISH was performed. FISH evidenced 4 disomic (1.6-2.0 balanced gene and chromosome 7) (16%) and 26 polysomic (84%) cases of which 7 were trisomic (2.2-3.0 balanced gene and

chromosome 7) and 19 were highly polysomic (3.1-4.4 balanced gene and chromosome Idasanutlin mouse 7) and 3 amplified (gene-to-chromosome 7 ratio ≥ 2). Sensitivity for CISH was 60%, specificity was 89%, the positive predictive value (PPV) was 50% and the negative Cell press predictive value (NPV) was 93% (Table 2). Table 2 Comparison between immunohistochemistry, CISH and FISH in 33 cell blocks from lung FNAC IHC score N° of cases CISH FISH     D T P A D T P A 0

9 1 5 3 0 2 2 5 0 1+ 3 1 0 2 0 0 0 3 0 2+ 5 1 0 4 0 0 0 5 0 3+ 16 2 7 6 2 2 5 6 3 Total 33 5 12 14 2 4 7 19 3 IHC: immunohistochemistry; CISH: chromogenic in situ hybridization; FISH: fluorescence in situ hybridization; D: disomy, 1.6-2.0 balanced gene and chromosome 7; T: trisomy, 2.2-3.0 balanced gene and chromosome 7; P: polysomy, 3.1-4.4 balanced gene and chromosome 7; A: amplified, gene-to-chromosome 7 ratio ≥2 Sensitivity 60%; Specificity 89%; Positive predictive value 50%; Negative predictive value 93% Table 3 reported the correlation between EGFR gene and chromosome 7 balanced polysomy by CISH and FISH. The overall concordance between FISH and CISH results was 97%. We observed 30 out of 33 cases not amplified (NA) and 2 NCSLC amplified (A) with both assays. CISH presented a gene-to-chromosome 7 ratio of 2.5 and 3 respectively and FISH a gene-to-chromosome 7 ratio of 2.8 and 3.3 respectively. Although there was a very low number of amplified cases, the 2 NSCLC FNAC with gene amplification by CISH were highly polysomic and this polysomy was confirmed by FISH.

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