Inhibitory effect of MicroRNA-136 targeting CD163 on the polarization of CD68+CD163+M2 macrophages

doi:10.12092/j.issn.1009-2501.2019.01.001

Abstract

Abstract:

AIM: To study the mechanism of MicroRNA (miR-136) inhibiting the polarization of CD68+M0 macrophages to CD68+CD163+M2 macrophages by targeting CD163. METHODS: The ratio of CD68+CD163+M2 macrophages in the tumor tissues and adjacent tissues of 20 patients with hepatocellular carcinoma was detected by flow cytometry, and the level of miR-136 in the tumor and adjacent tissues was detected by real-time fluorescence quantitative PCR (RT-qPCR). CD68+M0 macrophages were separated from the spleen, and the cells were divided into the control group (Control), the miRNA analogue group (miRNA mimic) and the miRNA inhibitor group (miRNA inhibitor). IL-4 and IL-13 10 ng/mL were used to induce M2 polarization. Western blot was used to detect the expression of CD163, TNF-α, iNOS and Arg1 in M2 macrophage. In the mechanism study, the levels of JAK1, STAT1 and STAT6 in the JAK/STAT signal were detected, and the expressions of PI3K and AKT1 in the PI3K/AKT signal were detected also. Luciferase reporter gene method was used to determine the target relationship between miR-136 and CD163. RESULTS: The proportion of M2 macrophages in HCC tissues was significantly higher than that of adjacent tissues, while the expression of miR-136 was significantly lower than that of adjacent tissues, and the expression of the two was negatively correlated. Luciferase reporter gene results showed that CD163 was the target gene of miR-136. The percentage of CD68+CD163+M2 macrophages in miRNA mimic was significantly lower than that in miRNA inhibitor, and there was no significant difference compared with control group. The low expression of JAK1, STAT6, PI3K, AKT1 in miRNA mimic showed that miR-136 could inhibit the expression of JAK1-STAT6 and PI3K-AKT1 to inhibit the polarization of M2 macrophage. CONCLUSION: miR-136 can target CD163 to inhibit the M2 polarization of macrophage, and its mechanism is related to the inhibition of JAK1-STAT6 and PI3K-AKT1, which is one of the regulatory mechanisms of M2 macrophages in the immunization of liver cancer.

Key words: MicroRNA-136, CD163, M2 macrophages, luciferase reporter genes

CLC Number:

R965.2
R730

HAN Chenyang, YANG Yi, LI Wenyang, WANG Jin, GUO Li. Inhibitory effect of MicroRNA-136 targeting CD163 on the polarization of CD68+CD163+M2 macrophages[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(1): 1-8.

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