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. 2009 May 19;100(12):1957–1965. doi: 10.1038/sj.bjc.6605060

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Copyright © 2009 Cancer Research UK

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Inhibition of Notch pathway prevents proliferation of osteosarcoma in vitro. Real-time PCR revealed four human osteosarcoma cell lines that express Notch2 (B). We carried out RT–PCR to determine which concentration of γ-Secretase inhibitor (GSI) X effectively inhibited Notch activity in osteosarcoma cells, and then measured the expression of the Notch pathway target HES1. γ-Secretase inhibitor X at 5 μM reduced mRNA levels of HES1 in 143B cells more than 60% (error bar means s.d.) (A). Western blot analysis showed that 5 μM GSI treatment reduced Notch2 intercellular domain (B). Growth of viable HOS and 143B cells over 3 days was slowed in dose-dependent fashion by GSI X (C). Real-time PCR revealed that siRNA knocked down CBF1 mRNA about 69% (D). Growth of HOS and 143B cells was slowed by CBF1 siRNA (E).