Skip to main content
Fig. 5 | Journal of Biomedical Science

Fig. 5

From: Tumor-associated macrophages promote resistance of hepatocellular carcinoma cells against sorafenib by activating CXCR2 signaling

Fig. 5

M2 TAMs release CXCL1 and CXCL2 to upregulate SOR resistance. A HepG2 cells, U937 cells, TAMsCM, and M2 MφsCyto were monocultured for 72 h, and HepG2 cells were cocultured with TAMsCM or M2 MφsCyto for 72 h. The CM from monocultures and cocultures were collected for measurement of CXCL1 and CXCL2 levels using ELISA assays. The bar graphs depict the relative protein expression. *P < 0.05; **P < 0.01; ***P < 0.001 vs. HepG2. B CXCL1 and CXCL2 mRNA expression in U937 cells, TAMsCM, M2 MφsCyto, and HepG2 cells without or with 72-h coculture was measured by qPCR. The bar graphs show the relative mRNA expression. ***P < 0.001, significant difference between groups. C HepG2 cells were monocultured and cocultured with TAMsCM or M2 MφsCyto for 72 h. The CM from monocultures and cocultures were collected to treat HepG2 cells in combination with SB225002 (50 μM) for 48 h followed by SOR (10 μM) for 48 h. The cells were collected to detect apoptosis using flow cytometry with double staining for Annexin V and PI. The bar graph shows the percentages of apoptotic cells. **P < 0.01; ***P < 0.001, significant difference between groups. D HepG2 cells were monocultured and cocultured with TAMsCM or M2 MφsCyto for 72 h. The CM from monocultures and cocultures were collected to treat HepG2 cells in combination with SB225002 (50 μM) for 48 h followed by CSC identification. The bar graph shows the percentages of CD44+/CD133+ cells measured by flow cytometry with double staining for CD44 and CD133. *P < 0.05; ***P < 0.001 vs. CM-HepG2. E After treatment with different CM as indicated in combination with SB225002 (50 μM) for 48 h, HepG2 cells were collected and seeded (2000 cells/well) in ultra-low attachment surface dishes for 14 days to form spheres. The bar graph shows the numbers of spheres with a diameter greater than 100 μm in each field. **P < 0.01; ***P < 0.001 vs. CM-HepG2. F HepG2 cells were treated with CXCL1 (10 ng/mL) or CXCL2 (50 ng/mL) in combination with SB225002 (50 μM) for 18 h followed by SOR (10 μM) for 48 h. Apoptosis was assessed by flow cytometry with double staining for Annexin V and PI. The bar graph depicts the percentages of apoptotic cells. **P < 0.01; ***P < 0.001, significant difference between groups. G HepG2 cells, Hep3B cells, and Huh7 cells were treated with CXCL1 (10 ng/mL) or CXCL2 (50 ng/mL) for 18 h, and then cells were collected for flow cytometry with double staining for CD44 and CD133. The bar graphs show the percentages of CD44+/CD133+ cells. *P < 0.05; ***P < 0.001 vs. control

Back to article page