Effect of gabexate mesilate and cetuximab, alone and in combination, on CRC cell viability.

<p>Evaluation by MTT assay of CACO-2, SW48, HT-29, Colo205, SW480, SW620, RKO, LS174T and HCT-116 cell viability after 24 and 48 h of treatment with cetuximab 100 µg/ml and gabexate mesilate 0.1–1 mM, alone and in combination. Three independent experiments were performed. For each cell line, the mean value of untreated samples was assumed as 100% and mean values of treated cells were plotted as percentages with respect to their matched controls. Cetux: cetuximab; GM: gabexate mesilate. *p<0.05; **p<0.001; ***p : not significant.</p

KRAS, BRAF and PIK3CA status of the CRC cell lines included in the study.

<p>KRAS, BRAF and PIK3CA status of the CRC cell lines included in the study.</p>*<p>from COSMIC database (<a href="http://www.sanger.ac.uk/genetics/CGP/cosmic/" target="_blank">http://www.sanger.ac.uk/genetics/CGP/cosmic/</a>).</p>**<p>KRAS/BRAF from COSMIC database; PIK3CA from Jhawer et al. 2008.</p

Effect of gabexate mesilate and cetuximab, alone and in combination, on CRC invasive potential.

<p>Evaluation by Boyden chamber invasion assay of CACO-2, SW48, HT-29, Colo205, SW480, SW620, RKO, LS174T and HCT-116 invasive potential after 6 hrs of treatment with cetuximab 100 µg/ml and gabexate mesilate 1 mM, alone and in combination. For each cell line, the mean value of untreated samples was assumed as 100% and mean values of treated cells were plotted as percentages with respect to their matched controls. Photographs of invading cells are representative of three independent experiments with similar findings. Cetux: cetuximab; GM: gabexate mesilate. Scale bar: 50 µm. *p<0.05; ***p : not significant.</p

Effect of gabexate mesilate and cetuximab, alone and in combination, on CRC conditioned medium-induced angiogenesis.

<p>Evaluation by in vitro Matrigel angiogenesis assay of EA.hy926 endothelial cell differentiation in capillary-like structures after 24 hrs incubation with the conditioned medium of CACO-2, SW48, HT-29, Colo205, SW480, SW620, RKO, LS174T and HCT-116 cells, previously treated for 6 hrs with cetuximab 100 µg/ml and gabexate mesilate 1 mM, alone and in combination. Tube formation index was assessed as described in Material and Methods section. For each cell line, the mean value of untreated samples was assumed as 100% and mean values of treated cells were plotted as percentages with respect to their matched controls. Photographs are representative of three independent experiments with similar findings. Cetux: cetuximab; GM: gabexate mesilate. Scale bar: 50 µm. *p<0.05; **p<0.001; ***p : not significant.</p

Additional file 3: Supplemental Figure S2. of MicroRNA profiling of primary pulmonary enteric adenocarcinoma in members from the same family reveals some similarities to pancreatic adenocarcinoma—a step towards personalized therapy

PCR analysis of miR-31*/-126*/-506/-508-3p/-514. Quantitative “miRNA-targeted” real-time PCR of five miRNA aberrantly expressed in the proband’s PEAC according to the microarray results showed that the microarray data were validated by PCR data as well as that the pattern of modulation of expression of these miRNA was similar in the proband’s brother. Columns and bars, average levels ± SD, compared using the 2-ΔΔCt method to a pool of normal lung tissues, dashed line. (PPTX 158 kb

Additional file 2: Supplemental Figure S1. of MicroRNA profiling of primary pulmonary enteric adenocarcinoma in members from the same family reveals some similarities to pancreatic adenocarcinoma—a step towards personalized therapy

Immunohistochemical analyses in the cytological sample of the proband’s brother. The stainings for TTF1, CK7, CDX-2, and CK20, summarized in the table, showed patterns of protein expression similar to the expression levels observed in the PEAC component of the proband’s sample. Left column (original magnification ×20); right column (original magnification ×40). (PPTX 32,747 kb

Additional file 1: of MicroRNA profiling of primary pulmonary enteric adenocarcinoma in members from the same family reveals some similarities to pancreatic adenocarcinoma—a step towards personalized therapy

Analyses of EGFR, KRAS, ALK and MSI. Detailed methods used for the analyses of EGFR, KRAS mutation, ALK rearrangement, MSI, and miRNA-PCR. (DOC 108 kb