Fig. 5: Experimental validation of the RSF model’s ability to predict chemotherapy efficiency in cell lines.

A, B NCI-H520, HCC15, and NCI-H226 cells were treated with vinorelbine or docetaxel in a dose-dependent manner. Cell viability was assessed to construct curve plots, after which the IC50 values were calculated (n = 3 per group; ∗∗, p < 0.01; ∗∗∗, p < 0.001; t test). C NCI-H520, HCC15, and NCI-H226 cells were plated into 6-well plates and then treated with DMSO, vinorelbine (3 μM), or docetaxel (10 μM). Fourteen days after implantation, the cells were stained with crystal violet and photographed with a camera (n = 5 per group). D NCI-H520, HCC15, and NCI-H226 cells were treated with DMSO, vinorelbine (3 μM), or docetaxel (10 μM). Following the colony formation assay, the number of colonies in each well was calculated by ImageJ (n = 5 per group). E NCI-H520, HCC15, and NCI-H226 cells were treated with DMSO or the TGF-β receptor I inhibitor SB525334 (5 μM) for 2 days. The cell lysates were immunoblotted with the indicated antibodies. F–G NCI-H520, HCC15, and NCI-H226 cells were treated with vinorelbine (3 μM) or docetaxel (10 μM) with either DMSO or SB525334 (5 μM) pretreatment. Cell viability was tested every day after cell implantation (n = 5 per group; ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001, ANOVA). H NCI-H520, HCC15, and NCI-H226 cells were treated with multiple doses of the TGF-β receptor I inhibitor SB525334 combined with vinorelbine or docetaxel for 2 days. Synergy plots depicting Gaddum’s noninteraction model (HSA) for NCI-H520, HCC15, and NCI-H226 cells were separately drawn. An HSA value that exceeds 10 indicates a synergistic interaction.