Cytotoxic Effects of Panobinostat and Romidepsin on Osteosarcoma Cells

SAOS2, SAOS2 LM7 and K7M2 cells were seeded 5 × 10³ cells per well in 96-well plates and in triplicate were treated with a dose range of panobinostat (Selleckchem, Houston, Texas, Cat #S1030; 1.8, 2.4, 3.3, 4.5, 6.0, 8.1, 11.0, 14.8 and 20 nM) or a dose range of romidepsin (Selleckchem Cat #S3020; 5.9, 8.8 13.2, 19.8, 29.6, 44.4, 66.7, 100 and 150 nM). For dual treatment with panobinostat and Carfilzomib (Selleck Cat #S2853) K7M2 and SAOS2 cells were plated at 5 × 10³ cells in 96-well plates and treated with panobinostat at a dose range (1.4, 1.8, 2.5, 3.3, 4.5, 6.1, 8.2, 11.1 and 15 nM) or Carfilzomib at a dose range (1.9, 2.6, 3.5, 4.7, 6.3, 8.5, 11.5, 15.6 and 21 nM) or combination of both drugs. IC₅₀ values were determined at 48 hours posttreatment using cell titer blue assay (Promega, Madison, Wisconsin, Cat # G8080). To determine the tubastatin (Selleckchem Cat # S2627), IC₅₀ value K7M2 cells were seeded at 2 × 10⁴ in 96-well plates and treated with the following concentrations (0, 1, 5, 10, 15, 100, 250, 500, 1250, 2500, 5000 and 10 000 nM). All treatments were done in triplicate and cell growth was assayed at 48 hours using MTS CellTiter 96 cell proliferation assay (Promega Cat # G5421). The impact of panobinostat and romidepsin on PDX derived cell lines was examined in a publicly available dataset (https://braid.stjude.org/masttour/).^{15 (link)}