Effects of halogenated boroxine on expression of human cancer drug targets in melanoma cell line

Recently it was found that dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH), is a potent and highly specific inhibitor of precancerous cell processes. We conducted gene expression profiling of human melanoma cells before and after treatment with two concentrations (0.1 and 1 mM) of this boron inorganic derivative in order to assess its effects on deregulation of genes associated with tumor pathways. Parallel trypan blue exclusion assay was performed to assess the cytotoxicity effects of this chemical. Treatment with K2(B3O3F4OH) induced a significant decrease of cell viability in melanoma cellline at both tested concentrations. Furthermore, these treatments caused deregulation of more than 30 genes known as common anti-tumor drug targets. IGF-1 and hTERT were found to be significantly downregulated and this result may imply potential use of K2(B3O3F4OH) as an inhibitor or human telomerase and insulin-like growth factor 1, both of which are associated with various tumor pathways. Upon the treatment of melanoma cells by K2(B3O3F4OH) in the above-mentioned manner, total RNA was extracted from harvested cells using RNA isolation kit (QIAGEN GmbH, Hilden, Germany).Human Cancer Drug Targets RT2 Profiler PCR method is used according to manufacturer’s instructions (RT2 Profiler PCR Array Handbook, Qiagen N. V., Venlo, Limburg, Netherlands) for a multigene profiling of expression of 84 genes (Table 2) on ABI 7300 real-time PCR apparatus (Applied Biosystems, Foster City, CA). The three cell populations used in the experiment (cell culture control sample and two cell cultures treated with K2(B3O3F4OH) in concentrations of 0.1mM and 1mM) were subsequently analyzed in three identical 96 wells array formats. The quality check up (false positive and false negative signals evaluation) was based on positive and negative controls included on arrays, as well as gDNA contamination probes.