As in the previous chapter to follow up and validate these preliminary data sets a more targeted approach using real time PCR was used with the same panel of rtPCR probes. This confirmed array findings of statistically significant increases in mRNA expression in response to IL-1ß in the canonical downstream targets of IL-1ß such as IL-6 and IL-1 alpha as seen in the literature (). It is known that cell type can have a huge impact on the signalling of BMPR2. In the cancer field, it has been demonstrated that inhibition of BMPR2 causes induction of apoptosis in chondrosarcoma cells () whereas in fibroblasts it has been shown to increase inflammation and metastasis (). I therefore decided to extend the panel of samples to include the aortic cells in rtPCR although I did not have array data to validate. I felt it was interesting enough to warrant investigation into this panel of genes in both cell types in the presence and absence of functional BMPR2. Literature notes that adhesion molecules VCAM and ICAM are increased upon inflammatory stimulus (, , ). I confirmed these effects and showed enhanced responses to IL-1ß when BMPR2 signalling is lost; in disease, this could go on to induce the macrophage infiltration and further inflammatory activation that causes the vascular damage (, ). These effects were interesting in their disease relevance and in the fact that the loss of BMPR2 does not cause these increases in cells derived from the systemic vascular bed and in fact in many cases the systemic cells become less inflammatory to IL-1ß when BMPR2 is non-functional. Adhesion molecules also are interesting as they play a major
As in the previous chapter to follow up and validate these preliminary data sets a more targeted approach using real time PCR was used with the same panel of rtPCR probes. This confirmed array findings of statistically significant increases in mRNA expression in response to IL-1ß in the canonical downstream targets of IL-1ß such as IL-6 and IL-1 alpha as seen in the literature (). It is known that cell type can have a huge impact on the signalling of BMPR2. In the cancer field, it has been demonstrated that inhibition of BMPR2 causes induction of apoptosis in chondrosarcoma cells () whereas in fibroblasts it has been shown to increase inflammation and metastasis (). I therefore decided to extend the panel of samples to include the aortic cells in rtPCR although I did not have array data to validate. I felt it was interesting enough to warrant investigation into this panel of genes in both cell types in the presence and absence of functional BMPR2. Literature notes that adhesion molecules VCAM and ICAM are increased upon inflammatory stimulus (, , ). I confirmed these effects and showed enhanced responses to IL-1ß when BMPR2 signalling is lost; in disease, this could go on to induce the macrophage infiltration and further inflammatory activation that causes the vascular damage (, ). These effects were interesting in their disease relevance and in the fact that the loss of BMPR2 does not cause these increases in cells derived from the systemic vascular bed and in fact in many cases the systemic cells become less inflammatory to IL-1ß when BMPR2 is non-functional. Adhesion molecules also are interesting as they play a major