Five variants that have been confirmed by Sanger sequencing, consisting of three frame shifting mutations at site 2430 and 2466 (one insertion and one deletion) and two missense mutations (C387W and L1600Q), were introduced into an existing NOTCH1 luciferase reporter construct and cotransfected into human bone osteosarcoma cells (U2OS) to determine relative activation of NOTCH1. Western blot was used to validate the change in sizes of the Notch1 proteins coded by two frameshift mutations. This evidence concerns the gene NOTCH1 and bone osteosarcoma.