Using oral cancer cell line UPCI: SCC078 to purify NuMA protein

Formation of multipolar spindles is closely linked to increased genomic instability and tumor progression. Centrosome hyperamplification is insufficient to initiate this mitotic defect, centrosome coalescence must be interrupted. Studies have indicated that cytoplasmic dynein is a key factor in preventing multipolarity, and overexpression of the NuMA protein is sufficient to mislocalize dynein from the spindle and abrogate the coalescence machinery. Because the mechanism by which NuMA can inhibit dynein is unclear, we are purifying NuMA to use in in vitro studies, to better understand how NuMA blocks dynein activity. Purifying NuMA from recombinant sources has not been successful; therefore we are utilizing a native source. We are using the oral cancer cell line UPCI:SCC078 as the source because it has nine copies of the NUMA1 gene. With modifications to the protocols used previously, our goal is to yield sufficient quantities of NuMA for biochemical analysis with purified NuMA.