As with other complex diseases, the genetic basis of osteoporosis is difficult to discover. Over the recent decade, several approaches have been used to discover genes affecting bone fragility, including the analysis of individuals or families with extreme osteoporotic phenotypes and, most notably, genome-wide association studies (GWAS).
In GWAS studies of loci associated with low mineral density, SNP polymorphisms in the FUBP3 and ANAPC1 genes have been identified, which have not yet been elucidated as to whether they affect bone remodeling. The purpose of the master's thesis was to functionally evaluate the role of both target genes during osteogenic differentiation. As part of the master's thesis, we introduced plasmid vectors into the osteosarcoma cells of HOS, which caused an increased or decreased gene expression for FUBP3 and ANAPC1 and observed how increased expression of FUBP3 and decreased expression of ANAPC1 affect the osteogenic differentiation of HOS cells. By measuring the degree of mineralization, we proved that increased expression of FUBP3 and silencing of ANAPC1 slow the osteogenic differentiation of HOS cells. We also nucleofected mesenchymal stem cells (MSCs) with the shANAPC1 plasmid and observed how the silencing of ANAPC1 affects the osteogenic differentiation of MSCs. Due to the failure of nucleofection of MSCs with the shANAPC1 plasmid, we could not define the role of silenced ANAPC1 expression in the osteogenic differentiation of MSCs. Using the qPCR method, we proved that increased expression of FUBP3 and decreased expression of ANAPC1 prevent the increase of osteoblastic markers OC, RUNX2, and COL1a1 during osteogenic differentiation. Thus, we additionally confirmed the influence of altered expression of FUBP3 and ANAPC1 genes on the osteogenic differentiation of HOS cells.
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