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. 2019 Nov 16:19:301.
doi: 10.1186/s12935-019-1018-4. eCollection 2019.

MicroRNA-140-5p inhibits salivary adenoid cystic carcinoma progression and metastasis via targeting survivin

Zhu Qiao  1 , Yue Zou  2 , Hu Zhao  1
Affiliations

MicroRNA-140-5p inhibits salivary adenoid cystic carcinoma progression and metastasis via targeting survivin

Zhu Qiao et al. Cancer Cell Int. .

Abstract

Background: Salivary adenoid cystic carcinoma (SACC) is one of the most frequent carcinomas derived from the salivary gland. Growing evidence implied the involvement of microRNAs (miRNAs) in SACC progression and metastasis. This study aimed to determine the regulatory role of miR-140-5p in SACC progression and metastasis and to explore the underlying mechanisms.

Materials and methods: MiR-140-5p and survivin mRNA expression levels were determined by quantitative real-time PCR; protein levels were evaluated by western blot assay; cell proliferation, growth, invasion, apoptosis and caspase-3 activity were evaluated by respective in vitro functional assays; xenograft nude mice model was used to assess the in vivo tumor growth; a luciferase reporter assay determined the interaction between miR-140-5p and survivin.

Results: MiR-140-5p overexpression suppressed SACC cell proliferation and invasion, induced cell apoptosis and inhibited in vivo tumor growth of SACC cells. The loss-of-function studies showed that miR-140-5p knockdown enhanced SACC cell proliferation and invasion, inhibited cell apoptosis and led to an accelerated in vivo tumor growth. The bioinformatics prediction and luciferase reporter assay revealed that miR-140-5p directly targeted survivin 3' untranslated region, and survivin was inversely regulated by miR-140-5p. Knockdown of survivin exerted tumor-suppressive effects on SACC cells, while enforced expression of survivin counteracted the tumor-suppressive actions of miR-140-5p overexpression in SACC cells. Mechanistically, miR-140-5p modulated the protein expression levels of apoptosis- and epithelial-mesenchymal transition-related mediators as well as matrix metallopeptidase-2/-9 via targeting survivin. More importantly, the down-regulation of miR-140-5p and the up-regulation of survivin were detected in the SACC clinical tissues, and miR-140-5 expression was inversely correlated with survivin mRNA expression level in SACC tissues.

Conclusion: Our data indicated that miR-140-5p suppressed SACC cell proliferation and invasion, induced cell apoptosis via regulating survivin expression. The present study provide evidence that that miR-140-5p could be a promising target for treating SACC, which requires further investigations.

Keywords: Apoptosis; Invasion; Proliferation; SACC; Survivin; miR-140-5p.

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Conflict of interest statement

Competing interestsThe authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
MiR-140-5p overexpression suppressed SACC-83 and SACC-LM cell proliferation, invasion and induced apoptosis, inhibited in vivo tumor growth. a qRT-PCR determined miR-140-5p expression levels in SACC-83 and SACC-LM cells after being transfected with mimics NC or miR-140-5p mimics (n = 3). b, c CCK-8 assay, d colony formation assay, e transwell invasion assay respectively determined cell proliferation, growth and invasion of SACC-83 and SACC-LM cells after being transfected with mimics NC or miR-140-5p mimics. f Flow cytometry and g caspase-3 activity assay respectively determine cell apoptotic rates and caspase-3 activity of SACC-83 and SACC-LM cells after being transfected with mimics NC or miR-140-5p mimics (n = 3). h In vivo tumor growth and (I) tumor weight from the xenograft nude mice inoculated with control SACC-LM cells or SACC-LM cells overexpressing miR-140-5p (n = 5). Significant difference between treatment groups were indicated as *P < 0.05, **P < 0.01 and ***P < 0.001
Fig. 2
Fig. 2
MiR-140-5p knockdown enhanced SACC-83 and SACC-LM cell proliferation, invasion and inhibited apoptosis, accelerated in vivo tumor growth. a qRT-PCR determined miR-140-5p expression levels in SACC-83 and SACC-LM cells after being transfected with inhibitors NC or miR-140-5p inhibitors (n = 3). b, c CCK-8 assay, d colony formation assay, e transwell invasion assay respectively determined cell proliferation, growth and invasion of SACC-83 and SACC-LM cells after being transfected with inhibitors NC or miR-140-5p inhibitors. f Flow cytometry and g caspase-3 activity assay respectively determine cell apoptotic rates and caspase-3 activity of SACC-83 and SACC-LM cells after being transfected with inhibitors NC or miR-140-5p inhibitors (n = 3). h In vivo tumor growth and i tumor weight from the xenograft nude mice inoculated with control SACC-LM cells or SACC-LM cells with miR-140-5p silencing (n = 5). Significant difference between treatment groups were indicated as *P < 0.05, **P < 0.01 and ***P < 0.001
Fig. 3
Fig. 3
MiR-140-5p targeted survivin 3′UTR and suppressed survivin expression. a Putative binding sites between miR-140-5p and survivin 3′UTR as predicted using online StarBase 2.0 tool. b, c Relative luciferase activity of different reporter constructs in SACC-83 and SACC-LM cells after being transfected with mimics NC or miR-140-5p mimics. d, e qRT-PCR and western blot assays respectively determined survivin mRNA and protein expression levels in SACC-83 and SACC-LM cells after being transfected with mimics NC or miR-140-5p mimics. f, g qRT-PCR and western blot assays respectively determined survivin mRNA and protein expression levels in SACC-83 and SACC-LM cells after being transfected with inhibitors NC or miR-140-5p inhibitors. N = 3. Significant difference between treatment groups were indicated as *P < 0.05 and **P < 0.01
Fig. 4
Fig. 4
MiR-140-5p regulated SACC-83 and SACC-LM cancer cell progression via targeting survivin. a, b CCK-8 assay, c colony formation assay, d transwell invasion assay respectively determined cell proliferation, growth and invasion of SACC-83 and SACC-LM cells after being treated with 100 nM YM-155 or normal medium (NC). e Flow cytometry and f caspase-3 activity assay respectively determine cell apoptotic rates and caspase-3 activity of SACC-83 and SACC-LM cells after being treated with 100 nM YM-155 or normal medium (NC). g qRT-PCR determined survivin mRNA expression levels in SACC-83 and SACC-LM cells after being transfected with pcDNA3.1 or pcDNA3.1-survivin. h, i CCK-8 assay, j colony formation assay, k transwell invasion assay respectively determined cell proliferation, growth and invasion of SACC-83 and SACC-LM cells after being co-transfected with mimics NC + pcDNA3.1, miR-140-5p mimics + pcDNA3.1 or miR-140-5p mimics + pcDNA3.1-survivin. f Flow cytometry and g caspase-3 activity assay respectively determine cell apoptotic rates and caspase-3 activity of SACC-83 and SACC-LM cells after being co-transfected with mimics NC + pcDNA3.1, miR-140-5p mimics + pcDNA3.1 or miR-140-5p mimics + pcDNA3.1-survivin. N = 3. Significant difference between treatment groups were indicated as *P < 0.05 and **P < 0.01
Fig. 5
Fig. 5
MiR-140-5p modulated the protein expression levels of apoptosis- and epithelial-mesenchymal transition-related mediators as well as matrix metallopeptidase-2/-9 via targeting survivin. Western blot assay determined protein expression levels of cleaved caspase-3/-9, XIAP, N-cadherin, vimentin, E-cadherin, matrix metallopeptidase-2/-9 in SACC-83 and SACC-LM cells after being co-transfected with mimics NC + pcDNA3.1, miR-140-5p mimics + pcDNA3.1 or miR-140-5p mimics + pcDNA3.1-survivin
Fig. 6
Fig. 6
Expression of miR-140-5p and survivin mRNA in SACC clinical samples. a, b qRT-PCR determined the miR-140-5p and survivin mRNA expression levels in SACC tissues and normal surround tissues from 35 patients with SACC. c Correlation analysis between miR-140-5p expression levels and survivin mRNA expression in SACC tissues. *P < 0.05 and **P < 0.01
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References

    1. Coca-Pelaz A, Rodrigo JP, Bradley PJ, Vander Poorten V, Triantafyllou A, Hunt JL, Strojan P, Rinaldo A, Haigentz M, Jr, Takes RP, et al. Adenoid cystic carcinoma of the head and neck—an update. Oral Oncol. 2015;51(7):652–661. doi: 10.1016/j.oraloncology.2015.04.005. - DOI - PubMed
    1. Gunduz AK, Yesiltas YS, Shields CL. Overview of benign and malignant lacrimal gland tumors. Curr Opin Ophthalmol. 2018;29(5):458–468. doi: 10.1097/ICU.0000000000000515. - DOI - PubMed
    1. Bradley PJ. Adenoid cystic carcinoma evaluation and management: progress with optimism! Curr Opin Otolaryngol Head Neck Surg. 2017;25(2):147–153. doi: 10.1097/MOO.0000000000000347. - DOI - PubMed
    1. Laurie SA, Ho AL, Fury MG, Sherman E, Pfister DG. Systemic therapy in the management of metastatic or locally recurrent adenoid cystic carcinoma of the salivary glands: a systematic review. Lancet Oncol. 2011;12(8):815–824. doi: 10.1016/S1470-2045(10)70245-X. - DOI - PubMed
    1. Andreasen S. Molecular features of adenoid cystic carcinoma with an emphasis on microRNA expression. APMIS. 2018;126(Suppl 140):7–57. doi: 10.1111/apm.12828. - DOI - PubMed