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MicroRNA-32 targeting PTEN enhances M2 macrophage polarization in the glioma microenvironment and further promotes the progression of glioma

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Abstract

This study was aimed to explore the molecular mechanism of macrophage polarization and its effect on glioma progression. THP1 cells were cocultured in conditioned medium from U87 human glioblastoma cells to simulate the glioma microenvironment. The expression of miR-32 and PTEN in THP1 cells was detected by real-time PCR. A luciferase reporter assay was conducted to confirm the target relation between miR-32 and PTEN. Western blot assays and ELISA were performed to detect PTEN, M2 macrophage-specific markers, PI3K/AKT signaling proteins, and apoptosis-related proteins. U87 cell proliferation was evaluated by CCK-8 and colony forming assays, and the migration ability of the cells was evaluated by Transwell and wound healing assays. The U87 culture supernatant promoted the M2 phenotype of THP1 cells. miR-32 was upregulated and PTEN was downregulated in THP1 cells with the M2 phenotype in the glioma microenvironment. Luciferase assays confirmed that PTEN expression was suppressed by miR-32 through interaction with the 3′UTR of PTEN. Overexpression of miR-32 suppressed PTEN expression in THP1 cells. Overexpression of miR-32 or downregulation of PTEN promoted the expression of M2 macrophage-specific markers, thereby enhancing M2 macrophage polarization. Additionally, miR-32 inhibited THP1 cell apoptosis via suppressing the PI3K/AKT signaling pathway. Most importantly, the proliferation and migration capacities of U87 cells treated with the THP1 culture supernatant after miR-32 overexpression were enhanced, and these effects could be reversed by cotransfection with pcDNA3.1-PTEN. miR-32 negatively modulates PTEN, thereby promoting M2 macrophage transformation through PI3K/AKT signaling, enhancing glioma proliferation and migration abilities.

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Authors and Affiliations

  1. Department of Neurosurgery, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121000, Liaoning, China

    Long Bao

  2. Department of Pediatrics, The First Affiliated Hospital of Jinzhou Medical University, No. 2, Section 5, People Street, Guta District, Jinzhou, 121000, Liaoning, China

    Xiang Li

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  1. Long Bao
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Correspondence to Xiang Li.

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Bao, L., Li, X. MicroRNA-32 targeting PTEN enhances M2 macrophage polarization in the glioma microenvironment and further promotes the progression of glioma. Mol Cell Biochem 460, 67–79 (2019). https://doi.org/10.1007/s11010-019-03571-2

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