Transfection of the TF gene and its effect on Bcl-2 expression and sensitivity to doxorubicin

, SK-N-SH cells lacking TF expression were transfected with the expression plasmid pcDNA3.1/TF and control plasmid (an empty neo pcDNA3.1 plasmid). Expression of transfected TF was detected by western blot assay. , the effect of interaction between TF and FVIIa on Bcl-2 and Bax expression. SK-N-SH transfected with TF and SH-EP1 expressing endogenous TF were both incubated with 10 nM FVIIa for the indicated time points. Cell extracts were then analyzed for Bcl-2 and Bax expression by western blot. Labels under Bcl-2 bands in the blot represent expression levels after normalization for actin, compared with untreated (0) samples (defined as 1 unit) , effect of enforced TF expression on cell survival of SK-N-SH cells in the presence or absence of FVIIa in response to doxorubicin. Cells transfected with TF and control cells (1 × 10) were treated with or without 10 nM FVIIa and different doses of doxorubicin. Cells were incubated for 48 h, then cell viability was determined by WST assay. , a similar WST assay of dose-dependent inhibition of FVIIa on doxorubicin-induced cell death. SK-N-SH cells transfected with TF were treated with 2 μM doxorubicin in the presence or absence of different concentrations of FVIIa as indicated.<p><b>Copyright information:</b></p><p>Taken from "Tissue factor/FVIIa activates Bcl-2 and prevents doxorubicin-induced apoptosis in neuroblastoma cells"</p><p>http://www.biomedcentral.com/1471-2407/8/69</p><p>BMC Cancer 2008;8():69-69.</p><p>Published online 6 Mar 2008</p><p>PMCID:PMC2275284.</p><p>