In Vitro Evaluation of Inhibitory Effect of Nuclear Factor-KappaB Activity by Small Interfering RNA on Pro-tumor Characteristics of M2-like Macrophages
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概要
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Tumor-associated macrophages (TAMs) have an alternatively activated macrophage phenotype (M2) and promote cancer cell proliferation, angiogenesis and metastasis. Nuclear factor-kappa B (NF-κB) is one of the master regulators of macrophage polarization. Here, we investigated the effect of inhibition of NF-κB activity by small interfering RNA (siRNA) on the pro-tumor response of macrophages located in the tumor microenvironment in vitro. We used mouse peritoneal macrophages cultured in conditioned medium from colon-26 cancer cells as an in vitro TAM model (M2-like macrophages). Transfection of NF-κB (p50) siRNA into M2-like macrophages resulted in a significant decrease in the secretion of interleukin (IL)-10 (a Th2 cytokine) and a significant increase of Th1 cytokine production (IL-12, tumor necrosis factor-α, and IL-6). Furthermore, vascular endothelial growth factor production and matrix metalloproteinase-9 mRNA expression in M2-like macrophages were suppressed by inhibition of NF-κB expression with NF-κB (p50) siRNA. In addition, there was a reduction of arginase mRNA expression and increase in nitric oxide production. The cytokine secretion profiles of macrophages cultured in conditioned medium from either B16BL6 or PAN-02 cancer cells were also converted from M2 to classically activated (M1) macrophages by transfection of NF-κB (p50) siRNA. These results suggest that inhibition of NF-κB activity in M2-like macrophages alters their phenotype toward M1.
著者
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HIGUCHI Yuriko
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University
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Kono Yusuke
Department Of Astronomy Graduate School Of Science The University Of Tokyo
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Okubo H.
Department Of Anatomy Shimane Medical University Institute For Immunology Faculty Of Medicine Kyoto University Department Of Dna Biology Tokai University School Of Medicine And Central Institute For Experimental Animals
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Hashida Mitsuru
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University
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Kawakami Shigeru
Graduate School of Biomedical Sciences, Nagasaki University
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Endo F.
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University
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