Synergistic Antitumour Properties of viscumTT in Alveolar Rhabdomyosarcoma
viscumTT inhibits cell proliferation and induces apoptosis synergistically in ARMS. (a) RMS-13 and RH-30 cells were incubated with viscum, TT, and viscumTT in increasing concentrations for 24 h. Cell proliferation was measured by CASY cell counter analysis, and the proliferation rate was calculated from the total cell numbers compared to untreated control cells. Apoptosis was assessed by annexin V/PI assay, and results were analysed by FlowJo Software. (b) RMS-13 cells were treated with viscum, TT, and viscumTT in increasing concentrations, and early cytotoxicity was measured by LDH assay. Results are presented as percentage of untreated control cells. (c) Apoptosis was confirmed by PARP cleavage. For this purpose, Western blot analyses were performed after treatment of RMS-13 cells with viscum, TT, and viscumTT. β-actin served as a loading control. Results are presented as means ± SD of three independent experiments. Webb’s fractional product (Fp) was calculated to assess synergism, values Fp > 1 display synergism. Mistletoe lectin (ML) and oleanolic acid (OA) concentrations were used as marker substances for viscum and TT, respectively.
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