Fig. 5: SIRT1 is involved in sorafenib-induced hyper-keratosis.
a HaCaT cells were transfected with non-targeting siRNA or siRNA targeting SIRT1. SIRT1 transcription level was detected by RT-qPCR (upper panel, n = 3) and the expression level of SIRT1 was determined by western blot (lower panel). b–e HaCaT cells were transfected with non-targeting siRNA or siRNA targeting SIRT1, followed by treatment with CdMCTRL or CdMSORA for 24 h (b, d) or treatment with or without s-HBEGF (2.5 ng/mL) for 24 h (c, e). The transcription levels of KRT1, KRT10, LORICRIN and IVL were measured by RT-qPCR (n = 3) (b, c). The cell survival rates were measured by SRB assay (n = 3) (d, e). f–i Mice were randomly divided into 4 groups. After injection of AAV1-shSirt1 adeno virus into the paws for 2 weeks, mice were treated with CMC-Na or sorafenib (100 mg/kg) daily by i.g. for 30 days (n = 5/group). f Representative western blot indicated the expression of SIRT1 in stratum corneum of mice in each group (n = 3/group). g Representative H&E staining and KRT5, KRT1, LORICRIN, SIRT1 immunohistochemistry staining were performed on the paws of mice. Scale bar, 50 μm. h Quantitative analysis of epidermal hyper-keratosis assessed by measuring the stratum corneum thickness. i s-HBEGF concentrations in the serum of each mouse were measured by ELISA (n = 5/group). Densitometric values are shown as optical density after GAPDH or ACTB normalization using Image J. Horizontal bars in (h) and (i) represent mean values. The results in (a), (b), (c), (d) and (e) are presented as the mean ± SD. Statistical analyses were performed using one-way ANOVA with LSD post hoc test in (b), (c), (h) and (i) and with Dunn’s post hoc test in (a). n.s. no significance; *P < 0.05; **P < 0.01; ***P < 0.001. SORA sorafenib, CdM HUVECs conditional medium, CTRL control.
