miR-217 inhibits laryngeal cancer metastasis by repressing AEG-1 and PD-L1 expression - PubMed (original) (raw)

. 2017 Jul 10;8(37):62143-62153.

doi: 10.18632/oncotarget.19121. eCollection 2017 Sep 22.

Xionghui Mao 1, Shu Zhao 2, Kaibin Song 1, Cheng Xiang 1, Yuanjing Lv 1, Huanyv Jiang 1, Lei Wang 1, Baojun Li 1, Xianguang Yang 1, Zhennan Yuan 1, Cheng Xiu 1, Hongxue Meng 3, Ji Sun 1

Affiliations

miR-217 inhibits laryngeal cancer metastasis by repressing AEG-1 and PD-L1 expression

Susheng Miao et al. Oncotarget. 2017.

Abstract

High incidences of laryngeal cancer have been reported recently. Increasing our understanding of the molecular mechanisms underlying this malignancy could reveal more effective approaches to treating laryngeal cancer patients and so improve their prognoses. In this study, we explored the biological effects of miR-217 on laryngeal cancer. miR-217 potently inhibited multiple metastatic traits, including cell migration, invasion, proliferation, apoptosis, and EMT, as well as angiogensis. These effects were achieved through downregulation of the miR-217 target gene, AEG-1 and PD-L1. Clinical expression and animal model studies further confirmed our results. These findings provide new insight into the physiological effects of miR-217 in laryngeal cancer and its potential therapeutic use.

Keywords: AEG-1; PD-L1; laryngeal cancer; metastasis; mir-217.

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Conflict of interest statement

CONFLICTS OF INTEREST No conflicts of interest to declare.

Figures

Figure 1

Figure 1. Expression of miR-217 in laryngeal cancer tissues

miR-217 levels were determined by qRT-PCR. (A) Box-plot of miR-217 expression patterns in carcinoma and paracarcinoma tissues (N=29, P < 0.0001). (B) Pairwise comparison of miR-217 expression between carcinoma and matching paracarcinoma tissues showing that miR-217 expression was reduced in 29/29 of the sample pairs.

Figure 2

Figure 2. miR-217 modulates metastatic traits in Hep2 cells

Overexpression of miR-217 in Hep2 cells resulted in (A) reduced migratory and invasive capability (N=5; scale bar, 50 μm), (B) reduced cell viability (N=3), and (C) cell cycle arrest (N=3).

Figure 3

Figure 3. miR-217 inhibits EMT in Hep2 cells

(A) Overexpression of miR-217 resulted in increased E-cadherin and decreased Vimentin in Hep2 cells. DAPI staining was used to detect nuclei. Scale bar, 50 μm. (B) Statistical histogram. N=3.

Figure 4

Figure 4. miR-217 inhibits angiogenesis

miR-217 inhibits (A) VEGF secretion in both Hep2 and HUVEC cells (N=3), and (B) tubule elongation and branching formation in HUVEC cells (N=3). Scale bar, 50 μm.

Figure 5

Figure 5. miR-217 directly regulates AEG-1 and PD-L1 in Hep2 cells

(A) Prediction of miR-217 binding sites on AEG-1 and PD-L1. (B) Prediction of miR-217 mutant binding sites on AEG-1 and PD-L1. (C) Luciferase activity in Hep2 cells infected with miR-217 or control after transfection with the indicated 3′-UTR-driven reporter plasmid or mutant 3′-UTR-driven reporter plasmid (N=3). (D) Real-time PCR results for endogenous AEG-1 and PD-L1 infected with miR-217 or NC (N=3). (E) Western blot results for endogenous AEG-1 and PD-L1 infected with miR-217 or NC.

Figure 6

Figure 6. AEG-1 activates downstream PD-L1

(A) qRT-PCR assay to measure PD-L1 mRNA levels when AEG-1 was overexpressed (N=3). (B) Western blot assay to measure PD-L1 protein levels when AEG-1 was overexpressed . (C) Luciferase assay showing activity for co-transfection of AEG-1 and promoter vector in Hep2 cells (N=3).

Figure 7

Figure 7. Immunohistochemistry detection of clinical human laryngeal carcinoma samples

The number of AEG1 or PD-L1 positive cells was counted in 10 randomly chosen fields for each patient (magnification: ×200). Score: left 3+ and right 1+.

Figure 8

Figure 8. Overexpression of miR-217 strongly inhibits metastasis in vivo

(A) Primary tumor growth upon subcutaneous implantation of 5 × 105 luciferase labeled Hep2 cells infected with the indicated vectors. The experiment was terminated after 6 weeks. (B) Statistical histogram (N=10).

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References

    1. Chu EA, Kim YJ. Laryngeal cancer: diagnosis and preoperative work-up. Otolaryngol Clin North Am. 2008;41:673–95. v. - PubMed
    1. Gupta GP, Massagué J. Cancer metastasis: building a framework. Cell. 2006;127:679–95. - PubMed
    1. Fidler IJ. The pathogenesis of cancer metastasis: the ‘seed and soil’ hypothesis revisited. Nat Rev Cancer. 2003;3:453–58. - PubMed
    1. Rothman KJ, Cann CI, Flanders D, Fried MP. Epidemiology of laryngeal cancer. Epidemiol Rev. 1980;2:195–209. - PubMed
    1. Siegel R, Naishadham D, Jemal A. Cancer statistics, 2013. CA Cancer J Clin. 2013;63:11–30. - PubMed

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