Knockdown of NLGN4X in melanoma cells

NLGN4X is a postsynaptic cell adhesion molecule that is important for synaptic function and has recently been implicated to play a role in various neuronal disorders. We detected high NLGN4X expression in non-metastatic melanoma cell lines and low NLGN4X expression in metastatic melanoma cells by Western blot and real time PCR analysis. Here, we used whole genome mRNA profiling to investigate the consequences of NLGN4X loss for melanoma progression. Therfore, we determined gene expression changes after NLGN4X knockdown and identified HIF1 pathway up-regulation in two non-metastatic melanoma cell lines. We transfected MCM1G and WM793B melanoma cell lines with either control non-targeting siRNA or NLGN4X siRNA for 24h (3 biological replicates per treatment and cell line). RNA was isolated 24h after knockdown, followed by hybridization on Affymetrix microarrays.