Fig. 3

The impact of sorafenib on IL4 secretion by HCC cells, and subsequent METs-mediated drug resistance. A The mRNA expression levels of macrophage activation associated cytokines in Hep3B cells after sorafenib treatment. n = 3. B ELISA detection of IL4 concentration in Hep1-6 and Hep3B culture media. NC: negative control. Sora: Hep1-6 and Hep3B cells were incubated with sorafenib. ELISA detection of the MPO-DNA concentration in the supernatant of M2-mBMDM and M2-THP-1 cells that were treated with C IL4 or D HCC cell media. n = 6 B–D. Blank: No treatment. NC: received treatment with IgG1 isotype and the media of Hepa1-6 or Hep3B. Sora: received treatment with IgG1 isotype and the media of Hepa1-6 or Hep3B treated by sorafenib. IL4-IN: received treatment with IL4 inhibitor (IL4-IN, neutralizing antibody) and the media of Hepa1-6 or Hep3B. Sora + IL4-IN: received treatment with IL4-IN and the media of Hepa1-6 or Hep3B treated by sorafenib. E Tumor photos and F tumor weight in Hepa1-6 model of HCC. n = 5. G Grouped tumor growth curve in Hepa1-6 model of HCC. H Tumor volume in Hepa1-6 model of HCC on day 25 after the first injection. n = 6. I Survival of Hepa1-6 model of HCC. n = 6. NC: negative control, treatment with IgG1 isotype. Sora: treatment with sorafenib and IgG1 isotype. IL4-IN: IL4 inhibition with neutralizing antibody. Sora + IL4-IN: treatment with sorafenib and IL4 neutralizing antibody. Statistical significance was calculated by two-tailed student’s t-test B, C, one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons test D, F and H, and Log-rank (Mantel-Cox) test I. Data represent the mean ± standard deviation (SD) of ≥ three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001