BACKGROUND:It has been generally demonstrated that the valine-to-glutamic acid substitution at position 600 (V600E) in the v-Raf murine sarcoma viral oncogene homolog B1 (BRAF) gene is an effective diagnostic=prognostic marker mainly for papillary thyroid carcinoma (PTC). The detection of this mutation typically has been achieved using DNA-based techniques. The recently introduced monoclonal V600E antibody (clone VE1) is likely to be an alternative strategy for detecting this mutation in thyroid lesions. The authors investigated molecular and immunocytohistochemical BRAF analyses in a prospective series of samples from patients with PTC. METHODS: Fifty-five prospective cytohistologic samples that were diagnosed as PTC were studied using both DNA BRAF testing and the VE1 antibody. The intensity of VE1 expression was graded from 0 (negative) to 31 (strong). RESULTS: All diagnoses were histologically confirmed. In total, 37 samples with mutated BRAF and 18 samples with wild-type BRAF were reported with 100% cytohistologic concordance. Cytologic VE1 expression revealed 25 negative samples (including 16 with 0 expression and 9 with 11 expression) and 30 positive samples (including 16 with 21 expression and 14 with 31 expression). On histology, there were 27 negative samples (20 with 0 expression and 7 with 11 expression) and 28 positive samples (14 with 21 expression and 14 with 31 expression). Four specimens with the BRAF mutation had discrepancies in VE1 intensity between cytology and histology. Furthermore, 6BRAF-mutated samples produced negative VE1 results. CONCLUSIONS: Although it has limitations, the VE1 antibody represents a feasible first-line approach for evaluating BRAF mutation status and may be a valid tool in the selection of samples for molecular analysis. The current report highlights the statistically significant difference between molecular and VE1 positivity in PTC (P <.0001). Nevertheless, in the authors' experience, BRAF mutations are more accurate for identifying VE1-negative cases.

H-thymidine incorporation. Expression of cytokine genes in colorectal cancer and autologous healthy mucosa was tested by quantitative, real-time PCR. A tumor microarray (TMA) including >1,200 colorectal cancer specimens was stained with GM-CSF-and M-CSF-specific antibodies. Clinicopathological features and overall survival were analyzed.Results: GM-CSF induced CD16 expression in 66% AE 8% of monocytes, as compared with 28% AE 1% in cells stimulated by M-CSF (P ¼ 0.011). GM-CSF but not M-CSF-stimulated macrophages significantly (P < 0.02) inhibited colorectal cancer cell proliferation. GM-CSF gene was expressed to significantly (n ¼ 45, P < 0.0001) higher extents in colorectal cancer than in healthy mucosa, whereas M-CSF gene expression was similar in healthy mucosa and colorectal cancer. Accordingly, IL1b and IL23 genes, typically expressed by M1 macrophages, were expressed to significantly (P < 0.001) higher extents in colorectal cancer than in healthy mucosa. TMA staining revealed that GM-CSF production by tumor cells is associated with lower T stage (P ¼ 0.02), "pushing" growth pattern (P ¼ 0.004) and significantly (P ¼ 0.0002) longer survival in mismatch-repair proficient colorectal cancer. Favorable prognostic effect of GM-CSF production by colorectal cancer cells was confirmed by multivariate analysis and was independent from CD16 þ and CD8 þ cell colorectal cancer infiltration. M-CSF expression had no significant prognostic relevance. Conclusions: GM-CSF production by tumor cells is an independent favorable prognostic factor in colorectal cancer. Clin Cancer Res; 20(12); 3094-106. Ó2014 AACR.