Long non‑coding RNA DILC is involved in sepsis by modulating the signaling pathway of the interleukin‑6/signal transducer and activator of transcription 3/Toll‑like receptor 4 axis

Huang,Weiping; Huang,Linqiang; Wen,Miaoyun; Fang,Ming; Deng,Yiyu; Zeng,Hongke

doi:10.3892/mmr.2018.9559

Long non‑coding RNA DILC is involved in sepsis by modulating the signaling pathway of the interleukin‑6/signal transducer and activator of transcription 3/Toll‑like receptor 4 axis

Authors:
- Weiping Huang
- Linqiang Huang
- Miaoyun Wen
- Ming Fang
- Yiyu Deng
- Hongke Zeng
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Affiliations: Department of Emergency and Critical Care Medicine, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510080, P.R. China
Published online on: October 16, 2018 https://doi.org/10.3892/mmr.2018.9559
Pages: 5775-5783

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Abstract

Sepsis is characterized by systemic inflammatory responses. In the present study, the role of deleted in liver cancer 1 (DILC), interleukin (IL)‑6, signal transducer and activator of transcription 3 (STAT3), and Toll‑like receptor 4 (TLR4) in the pathogenesis of sepsis was investigated. Reverse transcription‑quantitative polymerase chain reaction analysis and western blotting were performed to evaluate the effects of lipopolysaccharide (LPS) on the expression of DILC, IL‑6, STAT3, and TLR4, in addition to the effects of DILC and IL‑6 on the synthesis of tumor necrosis factor (TNF‑α), chemokine ligand 5 (CCL5), E‑selectin and C‑X‑C motif chemokine receptor 1 (CXCR1). In addition, the regulatory association between DILC, IL‑6, STAT3 and TLR4 was investigated. LPS reduced the expression level of DILC, and enhanced the expression of IL‑6, STAT3 and TLR4. DILC directly and negatively regulated the synthesis of IL‑6, as demonstrated by the markedly decreased luciferase activity in cells transfected with a wild‑type DILC plasmid. On the other hand, compared with the scramble control, DILC and IL‑6 small interfering (si)RNAs significantly suppressed the expression of IL‑6, STAT3 and TLR4. In addition, DILC siRNA enhanced the expression of IL‑6, STAT3 and TLR4, whereas the expression levels of TNF‑α, CCL5, E‑selectin and CXCR1 in LPS‑treated THP‑1 cells were downregulated following transfection with DILC and IL‑6 siRNAs. In patients with sepsis, DILC expression was inhibited, although the expression levels of IL‑6, STAT3 and TLR4 were upregulated. In addition, the expression levels of TNF‑α, CCL5, E‑selectin and CXCR1 in patients with sepsis were higher compared with normal subjects. Therefore, DILC may mediate the crosstalk between the cascades of IL‑6/STAT3 and TNF‑α signaling, indicating that DILC may act as a prognostic biomarker of sepsis, and may serve as a potential therapeutic target for the treatment of sepsis.

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1	Angus DC, Linde-Zwirble WT, Lidicker J, Clermont G, Carcillo J and Pinsky MR: Epidemiology of severe sepsis in the United States: Analysis of incidence, outcome, and associated costs of care. Crit Care Med. 29:1303–1310. 2001. View Article : Google Scholar : PubMed/NCBI
2	Bateman BT, Schmidt U, Berman MF and Bittner EA: Temporal trends in the epidemiology of severe postoperative sepsis after elective surgery: A large, nationwide sample. Anesthesiology. 112:917–925. 2010. View Article : Google Scholar : PubMed/NCBI
3	Hamano K, Gohra H, Noda H, Katoh T, Fujimura Y, Zempo N and Esato K: Increased serum interleukin-8: Correlation with poor prognosis in patients with postoperative multiple organ failure. World J Surg. 22:1077–1081. 1998. View Article : Google Scholar : PubMed/NCBI
4	Rubulotta FM, Ramsay G, Parker MM, Dellinger RP, Levy MM and Poeze M: Surviving Sepsis Campaign Steering Committee; European Society of Intensive Care Medicine; Society of Critical Care Medicine: An international survey: Public awareness and perception of sepsis. Crit Care Med. 37:167–170. 2009. View Article : Google Scholar : PubMed/NCBI
5	Kawamoto T, Ii M, Kitazaki T, Iizawa Y and Kimura H: Tak-242 selectively suppresses toll-like receptor 4-signaling mediated by the intracellular domain. Eur J Pharmacol. 584:40–48. 2008. View Article : Google Scholar : PubMed/NCBI
6	Biffl WL, Moore EE, Moore FA and Peterson VM: Interleukin-6 in the injured patient. Marker of injury or mediator of inflammation? Ann Surg. 224:647–664. 1996. View Article : Google Scholar : PubMed/NCBI
7	Pape HC, Schmidt RE, Rice J, van Griensven M, das Gupta R, Krettek C and Tscherne H: Biochemical changes after trauma and skeletal surgery of the lower extremity: Quantification of the operative burden. Crit Care Med. 28:3441–3418. 2000. View Article : Google Scholar : PubMed/NCBI
8	Giannoudis PV, Smith RM, Banks RE, Windsor AC, Dickson RA and Guillou PJ: Stimulation of inflammatory markers after blunt trauma. Br J Surg. 85:986–990. 1998. View Article : Google Scholar : PubMed/NCBI
9	Stensballe J, Christiansen M, Tønnesen E, Espersen K, Lippert FK and Rasmussen LS: The early IL-6 and IL-10 response in trauma is correlated with injury severity and mortality. Acta Anaesthesiol Scand. 53:515–521. 2009. View Article : Google Scholar : PubMed/NCBI
10	Adib-Conquy M and Cavaillon JM: Compensatory anti-inflammatory response syndrome. Thromb Haemost. 101:36–47. 2009. View Article : Google Scholar : PubMed/NCBI
11	Sander M, Irwin M, Sinha P, Naumann E, Kox WJ and Spies CD: Suppression of interleukin-6 to interleukin-10 ratio in chronic alcoholics: association with postoperative infections. Intensive Care Med. 28:285–292. 2002. View Article : Google Scholar : PubMed/NCBI
12	Mercer TR, Dinger ME and Mattick JS: Long non-coding RNAs: Insights into functions. Nat Rev Genet. 10:155–159. 2009. View Article : Google Scholar : PubMed/NCBI
13	Geisler S and Coller J: RNA in unexpected places: Long non-coding RNA functions in diverse cellular contexts. Nat Rev Mol Cell Biol. 14:699–712. 2013. View Article : Google Scholar : PubMed/NCBI
14	Ben-Neriah Y and Karin M: Inflammation meets cancer, with NF-kB as the matchmaker. Nat Immunol. 12:715–723. 2011. View Article : Google Scholar : PubMed/NCBI
15	Kagoya Y, Yoshimi A, Kataoka K, Nakagawa M, Kumano K, Arai S, Kobayashi H, Saito T, Iwakura Y and Kurokawa M: Positive feedback between NF-kB and TNF-α promotes leukemia-initiating cell capacity. J Clin Invest. 124:528–542. 2014. View Article : Google Scholar : PubMed/NCBI
16	Wang X, Sun W, Shen W, Xia M, Chen C, Xiang D, Ning B, Cui X, Li H, Li X, et al: Long non-coding RNA DILC regulates liver cancer stem cells via IL-6/STAT3 axis. J Hepatol. 64:1283–1294. 2016. View Article : Google Scholar : PubMed/NCBI
17	Greenhill CJ, Rose-John S, Lissilaa R, Ferlin W, Ernst M, Hertzog PJ, Mansell A and Jenkins BJ: IL-6 trans-signaling modulates TLR4-dependent inflammatory responses via STAT3. J Immunol. 186:1199–1208. 2011. View Article : Google Scholar : PubMed/NCBI
18	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
19	Li RJ, Gao CY, Guo C, Zhou MM, Luo J and Kong LY: The Anti-inflammatory Activities of two major withanolides from physalisminima via acting on NF-κB, STAT3, and HO-1 in LPS-stimulated RAW264.7 Cells. Inflammation. 40:401–413. 2017. View Article : Google Scholar : PubMed/NCBI
20	Schroeder A, Herrmann A, Cherryholmes G, Kowolik C, Buettner R, Pal S, Yu H, Müller-Newen G and Jove R: Loss of androgen receptor expression promotes a stem-like cell phenotype in prostate cancer through STAT3 signaling. Cancer Res. 74:1227–1237. 2014. View Article : Google Scholar : PubMed/NCBI
21	Korkaya H, Kim GI, Davis A, Malik F, Henry NL, Ithimakin S, Quraishi AA, Tawakkol N, D'Angelo R, Paulson AK, et al: Activation of an IL6 inflammatory loop mediates trastuzumab resistance in HER2+ breast cancer by expanding the cancer stem cell population. Mol Cell. 47:570–584. 2012. View Article : Google Scholar : PubMed/NCBI
22	Jinushi M, Chiba S, Yoshiyama H, Masutomi K, Kinoshita I, Dosaka-Akita H, Yagita H, Takaoka A and Tahara H: Tumor-associated macrophages regulate tumorigenicity and anticancer drug responses of cancer stem/initiating cells. Proc Natl Acad Sci USA. 108:12425–12430. 2011. View Article : Google Scholar : PubMed/NCBI
23	He G, Dhar D, Nakagawa H, Font-Burgada J, Ogata H, Jiang Y, Shalapour S, Seki E, Yost SE, Jepsen K, et al: Identification of liver cancer progenitors whose malignant progression depends on autocrine IL-6 signaling. Cell. 155:384–396. 2013. View Article : Google Scholar : PubMed/NCBI
24	Huang WL, Yeh HH, Lin CC, Lai WW, Chang JY, Chang WT and Su WC: Signal transducer and activator of transcription 3 activation up-regulates interleukin-6 autocrine production: A biochemical and genetic study of established cancer cell lines and clinical isolated human cancer cells. Mol Cancer. 9:3092010. View Article : Google Scholar : PubMed/NCBI
25	Judd LM, Bredin K, Kalantzis A, Jenkins BJ, Ernst M and Giraud AS: STAT3 activation regulates growth, inflammation, and vascularization in a mouse model of gastric tumorigenesis. Gastroenterology. 131:1073–1085. 2006. View Article : Google Scholar : PubMed/NCBI
26	Ogura H, Murakami M, Okuyama Y, Tsuruoka M, Kitabayashi C, Kanamoto M, Nishihara M, Iwakura Y and Hirano T: Interleukin-17 promotes autoimmunity by triggering a positive-feedback loop via interleukin-6 induction. Immunity. 29:628–636. 2008. View Article : Google Scholar : PubMed/NCBI
27	Sumimoto H, Imabayashi F, Iwata T and Kawakami Y: The BRAF-MAPK signaling pathway is essential for cancer-immune evasion in human melanoma cells. J Exp Med. 203:1651–1656. 2006. View Article : Google Scholar : PubMed/NCBI
28	Gao JW, Zhang AQ, Pan W, Yue CL, Zeng L, Gu W and Jiang J: Association between IL-6-174G/C polymorphism and the risk of sepsis and mortality: A systematic review and meta-analysis. PLoS One. 10:e01188432015. View Article : Google Scholar : PubMed/NCBI
29	Borden EC and Chin P: Interleukin-6: A cytokine with potential diagnostic and therapeutic roles. J Lab Clin Med. 123:824–829. 1994.PubMed/NCBI
30	Uusitalo-Seppälä R, Koskinen P, Leino A, Peuravuori H, Vahlberg T and Rintala EM: Early detection of severe sepsis in the emergency room: Diagnostic value of plasma C-reactive protein, procalcitonin, and interleukin-6. Scand J Infect Dis. 43:883–890. 2011. View Article : Google Scholar : PubMed/NCBI
31	Jin YH, Hou W, Kang HS, Koh CS and Kim BS: The role of interleukin-6 in the expression of PD-1 and PDL-1 on central nervous system cells following infection with Theiler's murine encephalomyelitis virus. J Virol. 87:11538–11551. 2013. View Article : Google Scholar : PubMed/NCBI
32	Takeda K and Akira S: Toll-like receptors in innate immunity. Int Immunol. 17:1–14. 2005. View Article : Google Scholar : PubMed/NCBI
33	Hu D, Wan L, Chen M, Caudle Y, LeSage G, Li Q and Yin D: Essential role of IL-10/STAT3 in chronic stress-induced immune suppression. Brain Behav Immun. 36:118–127. 2014. View Article : Google Scholar : PubMed/NCBI
34	Wittebole X, Coyle SM, Kumar A, Goshima M, Lowry SF and Calvano SE: Expression of tumour necrosis factor receptor and Toll-like receptor 2 and 4 on peripheral blood leucocytes of human volunteers after endotoxin challenge: A comparison of flow cytometric light scatter and immunofluorescence gating. Clin Exp Immunol. 141:99–106. 2005. View Article : Google Scholar : PubMed/NCBI
35	Härter L, Mica L, Stocker R, Trentz O and Keel M: Increased expression of toll-like receptor-2 and −4 on leukocytes from patients with sepsis. Shock. 22:403–409. 2004. View Article : Google Scholar : PubMed/NCBI
36	Hoebe K, Janssen E and Beutler B: The interface between innate and adaptive immunity. Nat Immunol. 5:971–974. 2004. View Article : Google Scholar : PubMed/NCBI
37	Kawai T and Akira S: The role of pattern-recognition receptors in innate immunity: Update on Toll-like receptors. Nat Immunol. 11:373–384. 2010. View Article : Google Scholar : PubMed/NCBI
38	Kumar V and Sharma A: Innate immunity in sepsis pathogenesis and its modulation: New immunomodulatory targets revealed. J Chemother. 20:672–683. 2008. View Article : Google Scholar : PubMed/NCBI
39	Weighardt H, Kaiser-Moore S, Schlautkötter S, Rossmann-Bloeck T, Schleicher U, Bogdan C and Holzmann B: Type I IFN modulates host defense and late hyperinflammation in septic peritonitis. J Immunol. 177:5623–5630. 2006. View Article : Google Scholar : PubMed/NCBI
40	Namath A and Patterson AJ: Genetic polymorphisms in sepsis. Crit Care Clin. 25(835–856): x2009.
41	Tsujimoto H, Ono S, Efron PA, Scumpia PO, Moldawer LL and Mochizuki H: Role of Toll-like receptors in the development of sepsis. Shock. 29:315–321. 2008.PubMed/NCBI
42	Zhang J, Yang J, Xu X, Liang L, Sun H, Liu G, Zhang L and Su Y: The influence of genetic polymorphisms in TLR4 and TIRAP, and their expression levels in peripheral blood, on susceptibility to sepsis. Exp Ther Med. 11:131–139. 2016. View Article : Google Scholar : PubMed/NCBI
43	Liu X, Su X, Xu S, Wang H, Han D, Li J, Huang M and Cao X: MicroRNA in vivo precipitation identifies miR-151-3p as a computational unpredictable miRNA to target Stat3 and inhibits innate IL-6 production. Cell Mol Immunol. 15:99–110. 2018. View Article : Google Scholar : PubMed/NCBI