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Identification of FBXW7α-regulated genes in M1-polarized macrophages in colorectal cancer by RNA sequencing.

Saudi Med J. 2019 Aug;40(8):766-773. doi:10.15537/smj.2019.8.24361
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摘要


OBJECTIVES:To determine the FBXW7α-regulated genes in tumor-polarized macrophages in colorectal cancer. METHODS:This experimental study was performed between June 2017 and March 2019. FBXW7α siRNA transfected RAW264.7 cells, together with the control group, were co-cultured with the colon cancer cell line, Colon-26. M1  marker production from the macrophages was determined by ELISA and quantitative reverse transcription-polymerase chain reaction. Whole genomic differential expression between the FBXW7α siRNA group and the control group were determined by RNA-sequencing analysis. The target site of the microRNA-205 gene was predicted using Targetscan and was verified by the luciferase assay. By transfecting mimics or inhibitors of microRNA-205, we explored the role of FBXW7α/microRNA-205 axis in regulating the polarization of tumor-associated macrophages (TAM). RESULTS:FBXW7α knockdown in RAW264.7 enhanced the expression of cyclooxigenase (COX)-2 and inducible nitric oxide synthase (iNOS), mRNA expression and IL6, IL12, p40, and tumor necrosis factor-α (TNFα) production upon co-culture with Colon-26 cells in vitro. Further, compared with the control group, 648 genes in total were enhanced and 416 targets were downregulated in FBXW7α siRNA transfected cells, among which miR-205 was the most significantly upregulated. SMAD1 was identified as an miR-205 target. The FBXW7α/miR-205 axis might regulate TAM polarization by affecting SMAD1 expression. CONCLUSION:These results prove that the FBXW7α/miR-205 axis plays an important role in TAM polarization and could facilitate further exploration of its molecular mechanism.

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