干扰TonEBP基因对Raw264.7细胞迁移、增殖和吞噬功能的影响

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Abstract Objective To investigate the effect of TonEBP gene on macrophage activity, migration, proliferation and phagocytosis. Methods The Raw264.7 cells transfected with TonEBP -shRNA lentivirus were screened by purinomycin to establish stable cell lines. The interference efficiency was evaluated by real-time PCR and Western blot. The lentivirus infection group with the highest interference efficiency and the empty lentivirus infection group were selected for subsequent experiments. Cells in each group were simultaneously cultured and incubated for 24 h.The cell activity was measured by CCK-8 assay, and the migration ability was observed by transwell migaration assay. Finally, the cell cycle and phagocytosis ability were detected by flow cytometry. Results The TonEBP stable cell line was successfully established and screened out. Compared with the wild type cell group, the interference efficiency of mRNA and protein was over 70% (P<0.05).Downregulation of TonEBP expression significantly reduced cell activity[24 h (0.39±0.02 vs 0.46±0.01),48 h (0.57±0.02 vs 0.63±0.02),72 h (0.71±0.02 vs 1.01±0.11),96 h (0.93±0.06 vs 1.42±0.04),120 h (1.26±0.06 vs 1.56±0.09) ] (P<0.05), red cell migration (88±7.00 vs 356±35.23), proliferation[ (38.73±2.57) ％ vs (50.20±2.29) ％] and phagocytosis [ (2.26±0.10) ％ vs (5.63±0.42) ％] (P<0.05). Conclusions Interference with TonEBP expression can effectively inhibit the activity of macrophages, reduce their migration, proliferation and phagocytosis ability. These results can contribute to further exploration of diseases related to macrophages.

Key words： tonicity-responsive enhancer binding protein Raw264.7 activity migration proliferation phagocytosis

Received: 10 August 2019

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	ZHU Zhe
	ZHAO Jihong
	YANG Guohong
	LI Tan
	HOU Yuehui
	LI Yuming

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ZHU Zhe,ZHAO Jihong,YANG Guohong等. Inhibitory effects of TonEBP gene silencing on activity, migration, proliferation and phagocytosis in Raw264.7 cells[J]. Med. J. Chin. Peop. Armed Poli. Forc., 2020, 31(2): 145-149.

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http://journal08.magtechjournal.com/Jwk_wjyx/EN/ OR http://journal08.magtechjournal.com/Jwk_wjyx/EN/Y2020/V31/I2/145

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