Figure 5

Overexpression of hCAP18 in MJ1105 cells increases metastasis formation in severe combined immunodeficiency (SCID) mice. (a) Metastasis formation displayed in a mouse injected with MJ1105-hCAP18 cells compared with control mouse treated with MJ1105 lacking hCAP18. The right panel shows tumour cells from ascites fluid detected by fluorescent transgene-coupled enhanced green fluorescent protein (eGFP) expression. (b) Expression analysis of hCAP18/L-37 in mouse tumours. (Left) Immunohistochemistry with anti-LL-37 antibodies demonstrating strong expression in transgenic tumours, and minor local induction in control tumours. Right, the expression of hCAP18 mRNA determined by reverse transcriptase PCR in tumours and transgenic cell lines. All comparisons are made to the expression of MJ1105 control cell line, which is set to one. (c) Western blot analysis demonstrates increased mitogen-activated protein kinase (MAPK) phosphorylation and increased degradation of phosphorylated ERBB2, in hCAP18 transgenic primary tumours compared with control primary tumours. On the left side of the blot, the control cell line is shown for comparison. Diagrams to the right illustrate the expression levels for respective tumour group, their statistic significance (two-tailed t-test, equal variance) indicated by asterix.