Analysis of the cytokine profiles in MC38CEA tumors and their cytotoxic effects on MC38CEA cells.

<p>(<b>A</b>) Relative protein and transcript levels of TNF, IFNγ and MCP-1 in lysates from wild type (WT), mock-transfected (M2) and ADAM17-silenced (S3) MC38CEA tumors measured by cytometric bead array immunoassay (CBA) or qRT-PCR, respectively. Average levels of each cytokine or its mRNA in M2 and/or WT tumor lysates were taken as 100% (CBA) or 1 (qRT-PCR). Control grups were as follows: M2 (three experiments, 5 mice per each experiment), WT (5 mice in one experiment) and both M2 (4 mice) and WT (5 mice) in one experiment. (<b>B</b>) Relative levels of TNF, IFNγ, and MCP-1 in sera from mice bearing WT, mock-transfected (M2), and ADAM17-silenced (S3) MC38CEA tumors at the end of the experiments. (<b>C, D</b>) The influence of TNF and IFNγ on MC38CEA viability. (<b>C</b>) Mock-transfected and ADAM17-silenced MC38CEA cells were incubated in DMEM/2% FCS with TNF (10 ng/ml) or IFNγ (50 ng/ml) or with TNF+IFNγ for 24, 48 or 72 h and then cell viability was assayed using MTT test. (<b>D</b>) Mock-transfected and ADAM17-silenced MC38CEA cells were incubated for 48 h in DMEM/2% FCS with TNF or IFNγ or with TNF+IFNγ at various concentrations and then cell viability was assayed using MTT test. (<b>E</b>) The influence of TNF and IFNγ on caspase activity in mock-transfected (M1 and M3) and ADAM17-silenced (S2 and S3) MC38CEA cells. Bars represent average values ± SD—for (<b>A</b>) and (<b>B</b>) from 5 independent experiments with at least 5 mice per each experimental group, * – <i>P</i><0.05 <i>vs.</i> M2 and WT controls; for (<b>C</b>) and (<b>D</b>) from 3 independent experiments performed in triplicates, * – <i>P</i><0.05 <i>vs.</i> untreated controls; (<b>E</b>) from 4 independent experiments performed in sextuplicates, * – <i>P</i><0.01 <i>vs.</i> untreated controls.