Influences of fibulin-5 and cleaved fibulin-5 on gene expression patterns of fibroblasts

To investigate the relevance and the impact of the cleaved FBLN5 variant on fibroblast’s physiology, we have employed whole genome microarrays. Human neonatal dermal fibroblasts from three different donors were seeded in duplicates on plates pre-coated with FBLN5, cleaved FBLN5 or heat denatured BSA as a control. After cultivation for 24 h, RNA was isolated and microarrays were performed. Statistical analysis of microarrays resulted in 367 genes significantly differentially expressed in cells cultured on a FBLN5 matrix and 525 genes differentially expressed in cells cultured on a cleaved FBLN5 matrix compared to control (adjusted p-value <0.05). To further validate these gene expression changes identified by microarray analysis 10 randomly choosen genes were reanalyzed by qRT-PCR (quantitative real time polymerase chain reaction). In nine cases the fold changes measured by qRT-PCR were comparable with those measured by microarray analysis, revealing reproducibility and validity of analysis. Fibulin-5 and cleaved FBLN5 contributed to changes in expression pattern of human neonatal dermal fibroblasts after cultivation for 24 h. Three different donors were used in technical duplicates.