Sunitinib suppresses M2 polarization of macrophages in tumor microenvironment to regulate hepatocellular carcinoma progression

Abstract

AbstractThis work aimed to investigate the role and mechanism of Sunitinib (Sun) in suppressing M2 polarization of macrophages in tumor microenvironment (TME). IL‐4 was applied to induce the M2 polarization of RAW264.7 cells, followed by treatment with Sun at 50 and 100 nM. Flow cytometry (FCM) was conducted to detect the proportion of F4/80 + CD206 + cells. Enzyme‐linked immunosorbent assay (ELISA) was performed to measure the levels of IL‐10, Arg‐1 and VEGF. Immunofluorescence (IF) staining was carried out to detect the expression of CD206 and Arg‐1. Besides, western‐Blot (WB) assay was performed to measure the levels of p‐JAK1 and p‐STAT6 proteins. After polarization, the macrophage culture medium was employed to culture hepatocellular carcinoma (HCC) Hca‐F cells. Thereafter, Transwell assays were conducted to examine cell invasion and migration, whereas plate clone formation assay was carried out to detect the clone forming capacity. In further experiments, cells were treated with the STAT6 inhibitor, or STAT6 inhibitor + Sun. Then, the polarization levels of RAW264.7 cells were detected. Moreover, this study established the xenograft tumor mouse model. Later, CD206 and Ki67 expression, IL‐10, Arg‐1 and VEGF expression levels in tissues, and p‐JAK1 and p‐STAT6 protein levels were detected by histochemical staining. Sun suppressed the M2 polarization of RAW264.7 cells. Compared with IL‐4 treatment, the proportion of F4/80 + CD206 + cells decreased. Meanwhile, the levels of IL‐10, Arg‐1 and VEGF were downregulated, and the phosphorylation level of JAK1‐STAT6 signaling was suppressed. After being cocultured with Hca‐F, the malignant behaviors of HCC cells were suppressed after Sun treatment. Similarly, STAT6 inhibitor treatment suppressed the M2 polarization, while the combined application of Sun did not further restrain the polarization level. In the mouse model, Sun suppressed the expression of CD206 and Ki67, simultaneously inhibiting the polarization of JAK1‐STAT6 signaling. Sunitinib can suppress the M2 polarization of macrophages to exert the anti‐HCC effect, which is its another anticancer mechanism