Fig 2: CCL2 promotes cervical cancer proliferation, migration, and invasion. (A,B) Flow cytometry results of phagocytosis assays in which HeLa or ME-180 cells were cocultured with or without SCs. The percentages of Ki+ CCR2+ cells are indicated in the upper right corner. (C–E) Cell migration and invasion assays were performed to assess the effect of RSC, CCL2, and CCR2 on cancer cells. The data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 and ns, not significant. (D,E) Refer to the statistical results.

Fig 3: Relationship between CCL2/CCR2 axis and PNI of cervical cancer in vivo. (A) Surgical images of the sciatic nerve. HeLa cells were injected into the left sciatic nerves. Mice treated with HeLa shCCR2 cells exhibited a smaller volume. The images to the right refer to the corresponding H&E staining results (50× magnification, scale bar, 200 µm). (B–D) Diameter of sciatic nerves, sciatic nerve function score and sciatic nerve function index of mice. (E) HeLa shNC cells injection resulted in the upregulation of N-cadherin and Snail. HeLa shCCR2 cells injection resulted in highly positive E-cadherin staining. *P < 0.05, ns, no significant. (200× magnification, scale bar, 100 µm).

Fig 4: Expression levels of CCL2/CCR2 and CXCL1/CXCR2 are correlated with HCC progression in the HCC model. Sprague Dawley rats were treated with diethylnitrosamine. After 8, 12 or 14 weeks, the rats were sacrificed to observe the development of HCC. The expression levels of CCL2, CCR2, CXCL1, and CXCR2 proteins were detected by immunohistochemical analysis. As CXCL8 is absent in rodents, we examined CXCL1, which is considered the functional equivalent of human CXCL8 in promoting neutrophil migration. The integral optical density (IOD) of the stained sections was calculated, and then Pearson’s correlation analysis was performed to investigate the relationship between the protein expression level and diethylnitrosamine treatment time. Pearson’s correlation analysis yielded the correlation coefficient (r) and the p-value. A Representative images of CCL2, CCR2, CXCL1, and CXCR2 immunostaining in the liver tissue of rats after 8, 12 and 14 weeks of diethylnitrosamine treatment. Asterisks indicate positive staining. Scale bar 200 µm. (B-E) Pearson's correlation analysis between B CCL2 and diethylnitrosamine treatment time (r = 0.70, p < 0.0001), C CCR2 and diethylnitrosamine treatment time (r = 0.74, p < 0.0001), D CXCL1 and diethylnitrosamine treatment time (r = 0.67, p < 0.0001), and E CCR2 and diethylnitrosamine treatment time (r = 0.71, p < 0.0001)

Fig 5: CCR2 and CXCR2 antagonists reduce the recruitment of TAMs/TANs in the primary HCC model. Sprague Dawley rats were treated with diethylnitrosamine for 14 weeks, then INCB334 and SCH527123 were administered via intraperitoneal injection. After 14 weeks, the rats were sacrificed and the livers were removed for immunohistochemical detection of CD68 and MPO. CD68 is a specific marker for TAMs while MPO is a specific marker for TANs. The expression levels of CD68 and MPO in the stained sections was assessed by IOD. A, C Representative images of CD68 and MPO immunostaining in livers from the indicated groups. Asterisks indicate positive staining. Scale bar, 200 µm. B Comparison of CD68 levels, based on IOD, of the indicated groups. D Comparison of MPO levels, based on IOD, of the indicated groups. Data are presented as mean ± SD. ns not statistically significant, *p < 0.05, **p < 0.01