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Suppression of miR-147b contributed to H37Rv-infected macrophage viability and migration in tuberculosis in vitro.

Microb Pathog. 2020 Jul;144:104125. Epub 2020 Mar 14
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摘要


BACKGROUND:Tuberculosis (TB) is a severe infectious disease. It was reported that microRNAs played important roles in tuberculosis. However, the role of miR-147b in the disease remained unveiling. METHODS:Tuberculosis cell model was established using macrophage THP-1 cells infected with H37Rv strain. RT-qPCR was first for examination of miR-147b relative expression. Cell viabilities were then measured with MTT. Cell transfection was to interfere the relative expression of miR-147b or C11orf87 in infected cells. RT-qPCR was adopted to confirm the transfection efficiency. Luciferase assay verified the binding sites between miR-147b and C11orf87. Migration was examined by scratch and relative protein expression of EMT biomarkers and phosphorylation of Pi3K and AKT were assessed via Western blot. RESULT:MiR-147b expression was higher and cell viability decreased in H32Rv-THP-1 cells. Cell viability was shown higher after miR-147b downregulation. Luciferase assay confirmed the binding. RT-qPCR found C11orf87 expression was lower in the H32Rv-THP-1 cells. MTT suggested that cell viability fell with the decrease of C11orf87 in infectious cells. Moreover, when H32Rv-THP-1 cells were co-transfected with miR-147b inhibitor and si-C11orf87, cell viability, migration and EMT and activation of Pi3K/AKT pathway was partially reversed compared with mere downregulation of miR-147b. CONCLUSION:miR-147b might regulate macrophage proliferation and migration through targeting C11orf87 via Pi3K/AKT pathway in Tuberculosis in vitro, which calls for in-depth inter-cellular researches and animal researches to further support that miR-147b/C11orf87 axis might be a potential therapeutic target for the molecular treatment of Tuberculosis in the future.

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