miR‑625‑5p suppresses inflammatory responses by targeting AKT2 in human bronchial epithelial cells

Qian,Fen-Hong; Deng,Xia; Zhuang,Qiong-Xin; Wei,Bin; Zheng,Dan-Dan

doi:10.3892/mmr.2019.9817

miR‑625‑5p suppresses inflammatory responses by targeting AKT2 in human bronchial epithelial cells

Authors:
- Fen-Hong Qian
- Xia Deng
- Qiong-Xin Zhuang
- Bin Wei
- Dan-Dan Zheng
View Affiliations

Affiliations: Department of Respiratory Medicine, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212000, P.R. China
Published online on: January 3, 2019 https://doi.org/10.3892/mmr.2019.9817
Pages: 1951-1957

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

Asthma is a common chronic inflammatory airway disease; however, whether microRNAs (miRs) could be used in the treatment of asthma remains unclear. The aim of the present study was to investigate the role of miR‑625‑5p in the inflammatory response of human bronchial epithelial cells (HBECs). Inflammation in the HBEC line, 16HBEC, was induced using different concentrations of lipopolysaccharide (LPS), which demonstrated that 1 µg/ml LPS was an appropriate concentration for further experiments. The association between protein kinase B2 (AKT2) and miR‑625‑5p was verified using a luciferase reporter assay. LPS was added to 16HBECs following the administration of miR‑625‑5p mimics or miR‑625‑5p inhibitors, and cells with silenced or overexpressed AKT2 levels. miR‑625‑5p was expressed at a high level in LPS‑activated 16HBECs. Overexpression of miR‑625‑5p inhibited interleukin (IL)‑6 and tumor necrosis factor (TNF)‑α secretion in 16HBECs. Inhibition of miR‑625‑5p enhanced LPS‑induced IL‑6 and TNF‑α secretion. miR‑625‑5p negatively regulated the expression of AKT2 in 16HBECs. A dual‑luciferase reporter assay system confirmed that miR‑625‑5p directly targeted the 3'untranslated region of AKT2. Transfection with a small interfering RNA against AKT2 inhibited inhibitor of κB phosphorylation. In brief, miR‑625‑5p may protect LPS‑induced HBECs by targeting AKT2 and inhibiting the nuclear factor‑κB signaling pathway. Therefore, miR‑625‑5p may function as an inhibitor of asthma airway inflammation in HBECs by targeting AKT2.

View Figures

View References

1	Schleimer RP, Kato A, Kern R, Kuperman D and Avila PC: Epithelium: At the interface of innate and adaptive immune responses. J Allergy Clin Immunol. 120:1279–1284. 2007. View Article : Google Scholar : PubMed/NCBI
2	Fang R, Cui Q, Sun J, Duan X, Ma X, Wang W, Cheng B, Liu Y, Hou Y and Bai G: PDK1/Akt/PDE4D axis identified as a target for asthma remedy synergistic with β2 AR agonists by a natural agent arctigenin. Allergy. 70:1622–1632. 2015. View Article : Google Scholar : PubMed/NCBI
3	Meister G and Tuschl T: Mechanisms of gene silencing by double-stranded RNA. Nature. 431:343–349. 2004. View Article : Google Scholar : PubMed/NCBI
4	Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, Sweet-Cordero A, Ebert BL, Mak RH, Ferrando AA, et al: MicroRNA expression profiles classify human cancers. Nature. 435:834–838. 2005. View Article : Google Scholar : PubMed/NCBI
5	Solberg OD, Ostrin EJ, Love MI, Peng JC, Bhakta NR, Hou L, Nguyen C, Solon M, Nguyen C, Barczak AJ, et al: Airway epithelial miRNA expression is altered in asthma. Am J RespirCrit Care Med. 186:965–974. 2012. View Article : Google Scholar
6	Martinez-Nunez RT, Bondanese VP, Louafi F, Francisco-Garcia AS, Rupani H, Bedke N, Holgate S, Howarth PH, Davies DE and Sanchez-Elsner T: A microRNA network dysregulated in asthma controls IL-6 production in bronchial epithelial cells. PLoS One. 9:e1116592014. View Article : Google Scholar : PubMed/NCBI
7	Matsukura S, Osakabe Y, Sekiguchi A, Inoue D, Kakiuchi Y, Funaki T, Yamazaki Y, Takayasu H, Tateno H, Kato E, et al: Overexpression of microRNA-155 suppresses chemokine expression induced by Interleukin-13 in BEAS-2B human bronchial epithelial cells. Allergol Int. 65 (Suppl):S17–S23. 2016. View Article : Google Scholar : PubMed/NCBI
8	Huo X, Zhang K, Yi L, Mo Y, Liang Y, Zhao J, Zhang Z, Xu Y and Zhen G: Decreased epithelial and plasma miR-181b-5p expression associates with airway eosinophilic inflammation in asthma. Clin Exp Allergy. 46:1281–1290. 2016. View Article : Google Scholar : PubMed/NCBI
9	Haj-Salem I, Fakhfakh R, Bérubé JC, Jacques E, Plante S, Simard MJ, Bossé Y and Chakir J: MicroRNA-19a enhances proliferation of bronchial epithelial cells by targeting TGFβR2 gene in severe asthma. Allergy. 70:212–219. 2015. View Article : Google Scholar : PubMed/NCBI
10	Rasmussen MH, Lyskjær I, Jersie-Christensen RR, Tarpgaard LS, Primdal-Bengtson B, Nielsen MM, Pedersen JS, Hansen TP, Hansen F, Olsen JV, et al: miR-625-3p regulates oxaliplatin resistance by targeting MAP2K6-p38 signalling in human colorectal adenocarcinoma cells. Nat Commun. 7:124362016. View Article : Google Scholar : PubMed/NCBI
11	Kirschner MB, Cheng YY, Badrian B, Kao SC, Creaney J, Edelman JJ, Armstrong NJ, Vallely MP, Musk AW, Robinson BW, et al: Increased circulating miR-625-3p: A potential biomarker for patients with malignant pleural mesothelioma. J Thorac Oncol. 7:1184–1191. 2012. View Article : Google Scholar : PubMed/NCBI
12	Zhou X, Zhang CZ, Lu SX, Chen GG, Li LZ, Liu LL, Yi C, Fu J, Hu W, Wen JM and Yun JP: miR-625 suppresses tumour migration and invasion by targeting IGF2BP1 in hepatocellular carcinoma. Oncogene. 34:965–977. 2015. View Article : Google Scholar : PubMed/NCBI
13	Zhang J, Zhang J, Zhang J, Qiu W, Xu S, Yu Q, Liu C, Wang Y, Lu A, Zhang J and Lu X: MicroRNA-625 inhibits the proliferation and increases the chemosensitivity of glioma by directly targeting AKT2. Am J Cancer Res. 7:1835–1849. 2017.PubMed/NCBI
14	Dong X, Xu M, Ren Z, Gu J, Lu M, Lu Q and Zhong N: Regulation of CBL and ESR1 expression by microRNA-22-3p, 513a-5p and 625-5p may impact the pathogenesis of dust mite-induced pediatric asthma. Int J Mol Med. 38:446–456. 2016. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Ozes ON, Mayo LD, Gustin JA, Pfeffer SR, Pfeffer LM and Donner DB: NF-kappaB activation by tumour necrosis factor requires the Akt serine-threonine kinase. Nature. 401:82–85. 1999. View Article : Google Scholar : PubMed/NCBI
17	Sizemore N, Lerner N, Dombrowski N, Sakurai H and Stark GR: Distinct roles of the Ikappa B kinase alpha and beta subunits in liberating nuclear factor kappa B (NF-kappa B) from Ikappa B and in phosphorylating the p65 subunit of NF-kappa B. J Biol Chem. 277:3863–3869. 2002. View Article : Google Scholar : PubMed/NCBI
18	Linnerth-Petrik NM, Santry LA, Moorehead R, Jücker M, Wootton SK and Petrik J: Akt isoform specific effects in ovarian cancer progression. Oncotarget. 7:74820–74833. 2016. View Article : Google Scholar : PubMed/NCBI
19	Riggio M, Perrone MC, Polo ML, Rodriguez MJ, May M, Abba M, Lanari C and Novaro V: AKT1 and AKT2 isoforms play distinct roles during breast cancer progression through the regulation of specific downstream proteins. Sci Rep. 7:442442017. View Article : Google Scholar : PubMed/NCBI
20	Wang Q, Yu WN, Chen X, Peng XD, Jeon SM, Birnbaum MJ, Guzman G and Hay N: Spontaneous hepatocellular carcinoma after the combined deletion of Akt isoforms. Cancer Cell. 29:523–535. 2016. View Article : Google Scholar : PubMed/NCBI
21	Attoub S, Arafat K, Hammadi NK, Mester J and Gaben AM: Akt2 knock-down reveals its contribution to human lung cancer cell proliferation, growth, motility, invasion and endothelial cell tube formation. Sci Rep. 5:127592015. View Article : Google Scholar : PubMed/NCBI
22	Kim CK, Nguyen TL, Lee SB, Park SB, Lee KH, Cho SW and Ahn JY: Akt2 and nucleophosmin/B23 function as an oncogenic unit in human lung cancer cells. Exp Cell Res. 317:966–975. 2011. View Article : Google Scholar : PubMed/NCBI
23	Lee MW, Kim DS, Lee JH, Lee BS, Lee SH, Jung HL, Sung KW, Kim HT, Yoo KH and Koo HH: Roles of AKT1 and AKT2 in non-small cell lung cancer cell survival, growth, and migration. Cancer Sci. 102:1822–1828. 2011. View Article : Google Scholar : PubMed/NCBI
24	Linnerth-Petrik NM, Santry LA, Petrik JJ and Wootton SK: Opposing functions of akt isoforms in lung tumor initiation and progression. PLoS One. 9:e945952014. View Article : Google Scholar : PubMed/NCBI
25	Hollander MC, Maier CR, Hobbs EA, Ashmore AR, Linnoila RI and Dennis PA: Akt1 deletion prevents lung tumorigenesis by mutant K-ras. Oncogene. 30:1812–1821. 2011. View Article : Google Scholar : PubMed/NCBI
26	Vergadi E, Vaporidi K, Theodorakis EE, Doxaki C, Lagoudaki E, Ieronymaki E, Alexaki VI, Helms M, Kondili E, Soennichsen B, et al: Akt2 deficiency protects from acute lung injury via alternative macrophage activation and miR-146a induction in mice. J Immunol. 192:394–406. 2014. View Article : Google Scholar : PubMed/NCBI
27	Gauna AE and Cha S: Akt2 deficiency as a therapeutic strategy protects against acute lung injury. Immunotherapy. 6:377–380. 2014. View Article : Google Scholar : PubMed/NCBI
28	Xu H, Sun Q, Lu L, Luo F, Zhou L, Liu J, Cao L, Wang Q, Xue J, Yang Q, et al: MicroRNA-218 acts by repressing TNFR1-mediated activation of NF-κB, which is involved in MUC5AC hyper-production and inflammation in smoking-induced bronchiolitis of COPD. Toxicol Lett. 280:171–180. 2017. View Article : Google Scholar : PubMed/NCBI
29	Luo F, Xu Y, Ling M, Zhao Y, Xu W, Liang X, Jiang R, Wang B, Bian Q and Liu Q: Arsenite evokes IL-6 secretion, autocrine regulation of STAT3 signaling, and miR-21 expression, processes involved in the EMT and malignant transformation of human bronchial epithelial cells. Toxicol Appl Pharmacol. 273:27–34. 2013. View Article : Google Scholar : PubMed/NCBI
30	Jardim MJ, Dailey L, Silbajoris R and Diaz-Sanchez D: Distinct microRNA expression in human airway cells of asthmatic donors identifies a novel asthma-associated gene. Am J Respir Cell Mol Biol. 47:536–542. 2012. View Article : Google Scholar : PubMed/NCBI
31	Gauthier M, Ray A and Wenzel SE: Evolving concepts of asthma. Am J Respir Crit Care Med. 192:660–668. 2015. View Article : Google Scholar : PubMed/NCBI