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. 2020 Feb 20:2020:3723781.
doi: 10.1155/2020/3723781. eCollection 2020.

MicroRNA-200a Promotes Phagocytosis of Macrophages and Suppresses Cell Proliferation, Migration, and Invasion in Nasopharyngeal Carcinoma by Targeting CD47

Yunteng Zhao  1 , Xiaoxiao Yu  1 , Haocheng Tang  1 , Ri Han  1 , Xianwen Wang  1 , Jianqi Wang  2 , Ke Wang  1 , Gang Li  1
Affiliations

MicroRNA-200a Promotes Phagocytosis of Macrophages and Suppresses Cell Proliferation, Migration, and Invasion in Nasopharyngeal Carcinoma by Targeting CD47

Yunteng Zhao et al. Biomed Res Int. .

Abstract

Nasopharyngeal carcinoma (NPC) causes severe oncogenic lesions in the nasopharynx. CD47, a transmembrane integrin-associated protein, plays a key role in the ability of tumor cells to escape phagocytosis, working as an immune checkpoint in the immune response. Besides this role, CD47 has been reported to regulate cell proliferation and migration. The present study addresses the relationship between CD47 and microRNA-200a and examines their regulatory mechanisms in NPC. Bioinformatics analyses and dual-luciferase reporter assays were used to confirm the putative relationship between miR-200a and CD47, and their interaction was further detected using western blotting and RT-PCR. Further, results showed that miR-200a affect NPC cell proliferation, migration, and invasion by regulating CD47. A cell phagocytosis assay showed that miR-200a and a CD47 monoclonal antibody increased the sensitivity of NPC cells to macrophage phagocytosis by inhibiting the functions of CD47. Additionally, miR-200a expression was suppressed and CD47 expression increased in both clinical NPC tissues and cell lines. Taken together, these results show the miR-200a/CD47 combination as a potential therapeutic for treatment of NPC.

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Conflict of interest statement

The authors have declared that no conflicts of interest exist.

Figures

Figure 1
Figure 1
Expression levels of miR-200a and CD47 in NPC. Notes: the expression of miR-200a was lower (a) and that of CD47 (b) was higher in NPC tissues than in nasopharyngitis samples. (c) Based on Spearman's correlation analysis, miR-200a expression was negatively correlated with CD47 levels. The expression of miR-200a was lower (d) while that of CD47 was higher (e) in NPC cells than in NP69 cells. ∗P < 0.05 vs. the NP69 group. Abbreviations: CD47: cluster of differentiation 47; NC: negative control.
Figure 2
Figure 2
miR-200a directly regulates CD47 expression in NPC cells. Notes: (a, b) based on a dual-luciferase assay, miR-200a was predicted to directly bind to the 3′UTR of CD47 mRNA, and cotransfection of the miR-200a mimic and wild-type promoter resulted in decreased luciferase activity. (c, d) At the mRNA level, CD47 expression was inhibited by the CD47 siRNA in both cell lines. (e, f) At the protein level, CD47 expression was inhibited by the miR-200a mimic in both cell lines. ∗P < 0.05 vs. the miR-NC group. ∗∗P < 0.05 vs. the miR-200a inhibitor group. Abbreviations: UTR: untranslated regions; WT: wild type; MUT: mutant-type; CD47: cluster of differentiation 47.
Figure 3
Figure 3
Proliferation and colony formation in CNE1 and CNE2 cells. Notes: (a, b) transfection of miR-200a and CD47 siRNA suppressed proliferation and (c, d) colony formation. Cotransfection with CD47 siRNA and miR-200a inhibitor rescued the effects of CD47 knockdown in CNE1 and CNE2 cells. ∗P < 0.05 vs. the miR-NC group. ∗∗P < 0.05 vs. the miR-200a inhibitor group. Abbreviations: CD47: cluster of differentiation 47; EdU: 5-ethynyl-2′-deoxyuridine; DAPI: 4′,6-diamidino-2-phenylindole; NC: negative control.
Figure 4
Figure 4
Migration and invasion of CNE1 and CNE2 cells. Notes: transfection with miR-200a and CD47 siRNA inhibited migration (a, c) and invasion (b, d) in CNE1 and CNE2 cells. Cotransfection with CD47 siRNA and miR-200a inhibitor rescued the effect of CD47 knockdown on the metastatic potential of CNE1 and CNE2 cells. ∗P < 0.05 vs. the miR-NC group. ∗∗P < 0.05 vs. the miR-200a inhibitor group. Abbreviations: CD47: cluster of differentiation 47; NC: negative control.
Figure 5
Figure 5
Anti-CD47 antibody and miR-200a promote phagocytosis of NPC cells by human macrophages. Notes: (a) human peripheral blood- (PB-) derived macrophages (green) are shown phagocytosing NPC cells (red) in the presence of a blocking anti-CD47 monoclonal antibody (B6H12) or IgG1 isotype control antibody. (b) Phagocytosed NPC cells (red) are shown in the miR-200a mimic and miR-NC groups. ∗P < 0.01. Abbreviations: CD47: cluster of differentiation 47; PBMC: peripheral blood mononuclear cell; IgG: immunoglobulin G.
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References

    1. Chua M. L. K., Wee J. T. S., Hui E. P., Chan A. T. C. Nasopharyngeal carcinoma. Lancet. 2016;387(10022):1012–1024. doi: 10.1016/S0140-6736(15)00055-0. - DOI - PubMed
    1. Petersson F. Nasopharyngeal carcinoma: a review. Seminars in Diagnostic Pathology. 2015;32(1):54–73. doi: 10.1053/j.semdp.2015.02.021. - DOI - PubMed
    1. Jiang F., Yu W., Zeng F., et al. PD-1 high expression predicts lower local disease control in stage IV M0 nasopharyngeal carcinoma. BMC Cancer. 2019;19(1):p. 503. doi: 10.1186/s12885-019-5689-y. - DOI - PMC - PubMed
    1. Yoshizaki T., Ito M., Murono S., Wakisaka N., Kondo S., Endo K. Current understanding and management of nasopharyngeal carcinoma. Auris Nasus Larynx. 2012;39(2):137–144. doi: 10.1016/j.anl.2011.02.012. - DOI - PubMed
    1. Strazzulla A., Barreca G. S., Giancotti A., et al. Nasopharyngeal carcinoma: review of the literature with a focus on therapeutical implications. Le Infezioni in Medicina. 2015;23(3):224–229. - PubMed