Fig 1: miR-520b inhibitor enhanced T cell activation in the tumor environment and guided the polarization of macrophages to M1, thereby inhibiting tumor growth. (a) The proportion of CD3, CD4, CD8, and NK1.1 cells. (b) The proportion of CD4+IFNγ and FOXP3 cells. (c) The proportion of CD206 and CD68 cells. The proportion of CD3, CD4, CD8, NK1.1, CD4+IFNγ, and CD68 cells increased, while FOXP3 and CD206 cells decreased in miR-520b inhibitor group compared with inhibitor NC group. However, the proportion of CD3, CD4, CD8, NK1.1, CD4+IFNγ, and CD68 cells decreased, while FOXP3 and CD206 cells increased after the addition of siPTEN. All experiments were repeated three times; #P < 0.05 vs the inhibitor NC group, and P < 0.05 vs the miR-520b inhibitor + siNC group. Differences between the two groups were analyzed using the student's t-test. One-way analysis of variance (ANOVA) was used for comparison between multiple groups.
Fig 2: The expression of miR-520b was high and PTEN was low in tumor tissues, and the immune microenvironment was changed. (a) miR-520b and PTEN expressions in tumor tissues were detected by qRT-PCR. Compared with tumor-adjacent tissues, miR-520b was highly expressed in breast tumor tissues, and PTEN was low expressed in breast tumor tissues. (b) miR-520b and PTEN expressions in breast cancer cells were detected by qRT-PCR. Compared with mammary epithelial cell line MCF-10A, miR-520b was highly expressed in breast cancer cell line MCF-7, and PTEN was low expressed in breast cancer cell line MCF-7. (c) CD45, CD3, CD4, CD8, and NK1.1 expressions in breast tumor tissues were detected by IF. CD45 immune cells were partially positive in breast tumor tissues, naive T cell marker CD3 was inhibited, and CD4, CD8, and NK1.1 were also inhibited. (d) IF detected CD206 and CD68 expression in breast tumor tissues. Compared with adjacent tissue, CD206 was strongly positive and CD68 was suppressed in breast tumor tissues. All experiments were repeated three times; ∗P < 0.05; scale bar = 25 μm; the magnification is 400 times. Differences between the two groups were analyzed using the student's t-test.
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