Knockdown of lncRNA ANRIL suppresses the production of inflammatory cytokines and mucin 5AC in nasal epithelial cells via the miR-15a-5p/JAK2 axis

Liu,Huo-Wang; Hu,Zhong-Liang; Li,Hao; Tan,Qi-Feng; Tong,Jing; Zhang,Yong-Quan

doi:10.3892/mmr.2020.11784

Knockdown of lncRNA ANRIL suppresses the production of inflammatory cytokines and mucin 5AC in nasal epithelial cells via the miR-15a-5p/JAK2 axis

Authors:
- Huo-Wang Liu
- Zhong-Liang Hu
- Hao Li
- Qi-Feng Tan
- Jing Tong
- Yong-Quan Zhang
View Affiliations

Affiliations: Department of Otolaryngology-Head and Neck Surgery, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China, Department of Pathology, Xiangya Hospital, Central South University, Changsha, Hunan 410007, P.R. China, Department of Otolaryngology-Head and Neck Surgery, The Fifth Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830000, P.R. China
Published online on: December 15, 2020 https://doi.org/10.3892/mmr.2020.11784
Article Number: 145
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The incidence of allergic rhinitis (AR) is increasing worldwide. Human nasal epithelial cells (HNECs) are the key cells in the occurrence of AR. Antisense non-coding RNA in the INK4 locus (ANRIL) was discovered to be involved in the progression of AR. However, the mechanism by which ANRIL mediates the progression of AR remains to be determined. The present study aimed to further explore the mechanism by which ANRIL regulates AR. Thereby, HNECs were treated with IL-13 to mimic AR in vitro. The mRNA expression levels of ANRIL, microRNA (miR)-15a-5p, JAK2, mucin 5AC (MUC5AC), granulocyte-macrophage colony-stimulating factor (GM-CSF) and eotaxin-1, and protein expression levels of JAK2, STAT3 and phosphorylated-STAT3 in HNECs were analyzed using reverse transcription-quantitative PCR and western blotting, respectively. ELISAs were used to detect the secretory levels of inflammatory cytokines and mucin in cell supernatants. In addition, a dual luciferase reporter assay was used to confirm the downstream target of ANRIL and the target gene of miR-15a-5p. The results revealed that the secretory levels of eotaxin-1, GM-CSF and MUC5AC were significantly upregulated by IL-13 in the supernatant of HNECs. The expression levels of ANRIL and JAK2 were also upregulated in IL-13-induced HNECs, while the expression levels of miR-15a-5p were downregulated. In addition, ANRIL was identified to bind to miR-15a-5p. The IL-13-induced upregulation of eotaxin-1, GM-CSF and MUC5AC mRNA expression and secretory levels was significantly inhibited by the genetic knockdown of ANRIL, while the miR-15a-5p inhibitor effectively reversed this effect. JAK2 was also discovered to be directly targeted by miR-15a-5p. The overexpression of JAK2 significantly suppressed the therapeutic effect of miR-15a-5p mimics on IL-13-induced inflammation in vitro. In conclusion, the findings of the present study suggested that the genetic knockdown of ANRIL may suppress the production of inflammatory cytokines and mucin in IL-13-treated HNECs via regulation of the miR-15a-5p/JAK2 axis. Thus, ANRIL may serve as a novel target for AR treatment.

View Figures

View References

1	Greiner AN, Hellings PW, Rotiroti G and Scadding GK: Allergic rhinitis. Lancet. 378:2112–2122. 2011. View Article : Google Scholar : PubMed/NCBI
2	Li L, Zhang L, Mo JH, Li YY, Xia JY, Bai XB, Xie PF, Liang JY, Yang ZF and Chen QY: Efficacy of indoor air purification in the treatment of Artemisia pollen-allergic rhinitis: A randomised, double-blind, clinical controlled trial. Clin Otolaryngol. 45:394–401. 2020. View Article : Google Scholar : PubMed/NCBI
3	Pazdro-Zastawny K, Kolator M, Krajewska J, Basiak-Rasała A, Górna S, Paluszkiewicz P, Zatoński M and Zatoński T: Lifestyle-related factors differentiating the prevalence of otorhinolaryngological diseases among 6–17-year-olds from Wrocław, Poland. Int J Pediatr Otorhinolaryngol. 132:1099342020. View Article : Google Scholar : PubMed/NCBI
4	Licari A, Castagnoli R, De Filippo M, Foiadelli T, Tosca MA, Marseglia GL and Ciprandi G: Current and emerging biologic therapies for allergic rhinitis and chronic rhinosinusitis. Expert Opin Biol Ther. 20:609–619. 2020. View Article : Google Scholar : PubMed/NCBI
5	Liu J, Zhang X, Zhao Y and Wang Y: The association between allergic rhinitis and sleep: A systematic review and meta-analysis of observational studies. PLoS One. 15:e02285332020. View Article : Google Scholar : PubMed/NCBI
6	Liang ZY, Deng YQ and Tao ZZ: A quantum dot-based lateral flow immunoassay for the rapid, quantitative, and sensitive detection of specific IgE for mite allergens in sera from patients with allergic rhinitis. Anal Bioanal Chem. 412:1785–1794. 2020. View Article : Google Scholar : PubMed/NCBI
7	Kenyon CC, Maltenfort MG, Hubbard RA, Schinasi LH, De Roos AJ, Henrickson SE, Bryant-Stephens TC and Forrest CB: Variability in diagnosed asthma in young children in a large pediatric primary care network. Acad Pediatr. 20:958–966. 2020. View Article : Google Scholar : PubMed/NCBI
8	Choi HS, Kim SL, Kim JH and Lee DS: The FDA-approved anti-asthma medicine ciclesonide inhibits lung cancer stem cells through Hedgehog signaling-mediated SOX2 regulation. Int J Mol Sci. 21:10142020. View Article : Google Scholar
9	Yi L, Cui Y, Xu Q and Jiang Y: Stabilization of LSD1 by deubiquitinating enzyme USP7 promotes glioblastoma cell tumorigenesis and metastasis through suppression of the p53 signaling pathway. Oncol Rep. 36:2935–2945. 2016. View Article : Google Scholar : PubMed/NCBI
10	Qin X, Zhu S, Chen Y, Chen D, Tu W and Zou H: Long non-coding RNA (lncRNA) CASC15 is upregulated in diabetes-induced chronic renal failure and regulates podocyte apoptosis. Med Sci Monit. 26:e9194152020. View Article : Google Scholar : PubMed/NCBI
11	Zhao J, Pu J, Hao B, Huang L, Chen J, Hong W, Zhou Y, Li B and Ran P: LncRNA RP11-86H7.1 promotes airway inflammation induced by TRAPM2.5 by acting as a ceRNA of miRNA-9-5p to regulate NFKB1 in HBECS. Sci Rep. 10:115872020. View Article : Google Scholar : PubMed/NCBI
12	Liu X, Zhu N, Zhang B and Xu SB: Long Noncoding RNA TCONS_00016406 attenuates lipopolysaccharide-induced acute kidney injury by regulating the miR-687/PTEN pathway. Front Physiol. 11:6222020. View Article : Google Scholar : PubMed/NCBI
13	Yang Y, Zhang Y, Yang Y, Guo J, Yang L, Li C and Song X: Differential expression of long noncoding RNAs and their function-related mRNAs in the peripheral blood of allergic rhinitis patients. Am J Rhinol Allergy. 34:508–518. 2020. View Article : Google Scholar : PubMed/NCBI
14	Zhu X, Wang X, Wang Y and Zhao Y: Exosomal long non-coding RNA GAS5 suppresses Th1 differentiation and promotes Th2 differentiation via downregulating EZH2 and T-bet in allergic rhinitis. Mol Immunol. 118:30–39. 2020. View Article : Google Scholar : PubMed/NCBI
15	Yue L, Yin X, Hao F, Dong J, Ren X, Xu O and Shan C: Long noncoding RNA linc00632 inhibits interleukin-13-induced inflammatory cytokine and mucus production in nasal epithelial cells. J Innate Immun. 12:116–128. 2020. View Article : Google Scholar : PubMed/NCBI
16	Qian X, Shi S and Zhang G: Long non-coding RNA antisense non-coding RNA in the INK4 locus expression correlates with increased disease risk, severity, and inflammation of allergic rhinitis. Medicine (Baltimore). 98:e152472019. View Article : Google Scholar : PubMed/NCBI
17	Li J, Xu X, Wei C, Liu L and Wang T: Long noncoding RNA NORAD regulates lung cancer cell proliferation, apoptosis, migration, and invasion by the miR-30a-5p/ADAM19 axis. Int J Clin Exp Pathol. 13:1–13. 2020.PubMed/NCBI
18	Yan Y, Peng Y, Ou Y and Jiang Y: MicroRNA-610 is downregulated in glioma cells, and inhibits proliferation and motility by directly targeting MDM2. Mol Med Rep. 14:2657–2664. 2016. View Article : Google Scholar : PubMed/NCBI
19	Chickooree D, Zhu K, Ram V, Wu HJ, He ZJ and Zhang S: A preliminary microarray assay of the miRNA expression signatures in buccal mucosa of oral submucous fibrosis patients. J Oral Pathol Med. 45:691–697. 2016. View Article : Google Scholar : PubMed/NCBI
20	Wang H, Guo Y, Mi N and Zhou L: miR-101-3p and miR-199b-5p promote cell apoptosis in oral cancer by targeting BICC1. Mol Cell Probes. 52:1015672020. View Article : Google Scholar : PubMed/NCBI
21	Hawkins LJ and Storey KB: MicroRNA expression in the heart of Xenopus laevis facilitates metabolic adaptation to dehydration. Genomics. 112:3525–3536. 2020. View Article : Google Scholar : PubMed/NCBI
22	Yang X, Zhu X, Yan Z, Li C, Zhao H, Ma L, Zhang D, Liu J, Liu Z, Du N, et al: miR-489-3p/SIX1 axis regulates melanoma proliferation and glycolytic potential. Mol Ther Oncolytics. 16:30–40. 2019. View Article : Google Scholar : PubMed/NCBI
23	Chen J, Wang J, Li H, Wang S, Xiang X and Zhang D: p53 activates miR-192-5p to mediate vancomycin induced AKI. Sci Rep. 6:388682016. View Article : Google Scholar : PubMed/NCBI
24	Shen ED, Liu B, Yu XS, Xiang ZF and Huang HY: The effects of miR-1207-5p expression in peripheral blood on cisplatin-based chemosensitivity of primary gallbladder carcinoma. OncoTargets Ther. 9:3633–3642. 2016. View Article : Google Scholar
25	Vakkilainen S, Mäkitie R, Klemetti P, Valta H, Taskinen M, Husebye ES and Mäkitie O: A wide spectrum of autoimmune manifestations and other symptoms suggesting immune dysregulation in patients with cartilage-hair hypoplasia. Front Immunol. 9:24682018. View Article : Google Scholar : PubMed/NCBI
26	Ma Z, Teng Y, Liu X, Li J, Mo J, Sha M and Li Y: Identification and functional profiling of differentially expressed long non-coding RNAs in nasal mucosa with allergic rhinitis. Tohoku J Exp Med. 242:143–150. 2017. View Article : Google Scholar : PubMed/NCBI
27	Liu HC, Liao Y and Liu CQ: miR-487b mitigates allergic rhinitis through inhibition of the IL-33/ST2 signaling pathway. Eur Rev Med Pharmacol Sci. 22:8076–8083. 2018.PubMed/NCBI
28	Wang L, Lv Q, Song X, Jiang K and Zhang J: ADRB2 suppresses IL-13-induced allergic rhinitis inflammatory cytokine regulated by miR-15a-5p. Hum Cell. 32:306–315. 2019. View Article : Google Scholar : PubMed/NCBI
29	Zhu M, Yang M, Yang Q, Liu W, Geng H, Pan L, Wang L, Ge R, Ji L, Cui S, et al: Chronic hypoxia-induced microvessel proliferation and basal membrane degradation in the bone marrow of rats regulated through the IL-6/JAK2/STAT3/MMP-9 pathway. BioMed Res Int. 2020:92047082020.PubMed/NCBI
30	Wang T, Chen D, Wang P, Xu Z and Li Y: miR-375 prevents nasal mucosa cells from apoptosis and ameliorates allergic rhinitis via inhibiting JAK2/STAT3 pathway. Biomed Pharmacother. 103:621–627. 2018. View Article : Google Scholar : PubMed/NCBI
31	Tang H, Wang H, Bai J, Ding M, Liu W, Xia W, Luo Q, Xu G, Li H and Fang J; Nasal Health Group China, : Sensitivity of MUC5AC to topical corticosteroid is negatively associated with interleukin-17A in patients with allergic rhinitis. Am J Rhinol Allergy. 26:359–364. 2012. View Article : Google Scholar : PubMed/NCBI
32	Zhai KF, Zheng JR, Tang YM, Li F, Lv YN, Zhang YY, Gao Z, Qi J, Yu BY and Kou JP: The saponin D39 blocks dissociation of non-muscular myosin heavy chain IIA from TNF receptor 2, suppressing tissue factor expression and venous thrombosis. Br J Pharmacol. 174:2818–2831. 2017. View Article : Google Scholar : PubMed/NCBI
33	Zhai KF, Duan H, Cui CY, Cao YY, Si JL, Yang HJ, Wang YC, Cao WG, Gao GZ and Wei ZJ: Liquiritin from glycyrrhiza uralensis attenuating rheumatoid arthritis via reducing inflammation, suppressing angiogenesis, and inhibiting MAPK signaling pathway. J Agric Food Chem. 67:2856–2864. 2019. View Article : Google Scholar : PubMed/NCBI
34	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−∆∆C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
35	Zhai KF, Duan H, Chen Y, Khan GJ, Cao WG, Gao GZ, Shan LL and Wei ZJ: Apoptosis effects of imperatorin on synoviocytes in rheumatoid arthritis through mitochondrial/caspase-mediated pathways. Food Funct. 9:2070–2079. 2018. View Article : Google Scholar : PubMed/NCBI
36	Ma Y, Shi L and Zheng C: Microarray analysis of lncRNA and mRNA expression profiles in mice with allergic rhinitis. Int J Pediatr Otorhinolaryngol. 104:58–65. 2018. View Article : Google Scholar : PubMed/NCBI
37	Teng Y, Zhang R, Liu C, Zhou L, Wang H, Zhuang W, Huang Y and Hong Z: miR-143 inhibits interleukin-13-induced inflammatory cytokine and mucus production in nasal epithelial cells from allergic rhinitis patients by targeting IL13Rα1. Biochem Biophys Res Commun. 457:58–64. 2015. View Article : Google Scholar : PubMed/NCBI
38	Incorvaia C, Cavaliere C, Frati F and Masieri S: Allergic rhinitis. J Biol Regul Homeost Agents. 32 (Suppl 1):61–66. 2018.PubMed/NCBI
39	Zhou B, Li L, Qiu X, Wu J, Xu L and Shao W: Long non-coding RNA ANRIL knockdown suppresses apoptosis and pro-inflammatory cytokines while enhancing neurite outgrowth via binding microRNA-125a in a cellular model of Alzheimer's disease. Mol Med Rep. 22:1489–1497. 2020. View Article : Google Scholar : PubMed/NCBI
40	Ping Y, Zhou Y, Hu J, Pang L, Xu C and Xiao Y: Dissecting the functional mechanisms of somatic copy-number alterations based on dysregulated ceRNA networks across cancers. Mol Ther Nucleic Acids. 21:464–479. 2020. View Article : Google Scholar : PubMed/NCBI
41	Wu H, Tian X and Zhu C: Knockdown of lncRNA PVT1 inhibits prostate cancer progression in vitro and in vivo by the suppression of KIF23 through stimulating miR-15a-5p. Cancer Cell Int. 20:2832020. View Article : Google Scholar : PubMed/NCBI
42	Zhu Y, Zhang X, Wang L, Zhu X, Xia Z, Xu L and Xu J: FENDRR suppresses cervical cancer proliferation and invasion by targeting miR-15a/b-5p and regulating TUBA1A expression. Cancer Cell Int. 20:1522020. View Article : Google Scholar : PubMed/NCBI
43	Lou Y and Huang Z: microRNA-15a-5p participates in sepsis by regulating the inflammatory response of macrophages and targeting TNIP2. Exp Ther Med. 19:3060–3068. 2020.PubMed/NCBI
44	Yu L, Liu Z, He W, Chen H, Lai Z, Duan Y, Cao X, Tao J, Xu C, Zhang Q, et al: Hydroxysafflor yellow A confers neuroprotection from focal cerebral ischemia by modulating the crosstalk between JAK2/STAT3 and SOCS3 signaling pathways. Cell Mol Neurobiol. 40:1271–1281. 2020. View Article : Google Scholar : PubMed/NCBI
45	Fang XY, Zhang H, Zhao L, Tan S, Ren QC, Wang L and Shen XF: Corrigendum to ‘A new xanthatin analogue 1β-hydroxyl-5α-chloro-8-epi-xanthatin induces apoptosis through ROS-mediated ERK/p38 MAPK activation and JAK2/STAT3 inhibition in human hepatocellular carcinoma’ [Biochimie 152C (2018) 43–52]. Biochimie. 171-172:21–22. 2020. View Article : Google Scholar : PubMed/NCBI
46	Wang J, Du A, Wang H and Li Y: MiR-599 regulates LPS-mediated apoptosis and inflammatory responses through the JAK2/STAT3 signaling pathway via targeting ROCK1 in human umbilical vein endothelial cells. Clin Exp Pharmacol Physiol. 47:1420–1428. 2020. View Article : Google Scholar : PubMed/NCBI
47	Peng DH, Liu YY, Chen W, Hu HN and Luo Y: Epidermal growth factor alleviates cerebral ischemia-induced brain injury by regulating expression of neutrophil gelatinase-associated lipocalin. Biochem Biophys Res Commun. 524:963–969. 2020. View Article : Google Scholar : PubMed/NCBI
48	Kabiri M, Hemmatpour A, Zare F, Hadinedoushan H and Karimollah A: Paroxetine modulates immune responses by activating a JAK2/STAT3 signaling pathway. J Biochem Mol Toxicol. 34:e224642020. View Article : Google Scholar : PubMed/NCBI