IGF2BP2 knockdown inhibits LPS‑induced pyroptosis in BEAS‑2B cells by targeting caspase 4, a crucial molecule of the non‑canonical pyroptosis pathway

Wang,Jing; Yuan,Xiaoli; Ding,Ning

doi:10.3892/etm.2021.10025

IGF2BP2 knockdown inhibits LPS‑induced pyroptosis in BEAS‑2B cells by targeting caspase 4, a crucial molecule of the non‑canonical pyroptosis pathway

Authors:
- Jing Wang
- Xiaoli Yuan
- Ning Ding
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Affiliations: Department of Emergency Medicine, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, P.R. China
Published online on: April 8, 2021 https://doi.org/10.3892/etm.2021.10025
Article Number: 593
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Insulin‑like growth factor 2 (IGF2) mRNA‑binding protein 2 (IGF2BP2) is a secreted protein that can bind to IGF2 and has been reported to promote inflammation. The data from the ENCORI database have predicted that IGF2BP2 can bind caspase 4, which mediates pyroptosis and promotes airway inflammation and lipopolysaccharide (LPS)‑induced lung injury. The present study investigated whether IGF2BP2 can regulate LPS‑induced lung cell inflammation by targeting caspase 4. Therefore, the non‑tumorigenic lung epithelial cell line Beas‑2B was transfected with short hairpin RNA (shRNA)‑IGF2BP2 and stimulated with LPS. A number of parameters, including cell viability, production of interleukin (IL)‑1β and IL‑18, activation of gasdermin D (GSDMD) and the expression levels of IGF2BP2, caspase 4 and cleaved‑caspase 1, were subsequently assessed using CCK‑8, ELISA kits, western blotting and immunofluorescence staining, respectively. RNA pull‑down assay was used to probe the possible interaction between IGF2BP2 and caspase 4 RNA. LPS treatment was found to inhibit cell viability, trigger IL‑1β and IL‑18 production and increase IGF2BP2 expression in a concentration‑dependent manner. Compared with cells transfected with shRNA‑negative control, cells that were transfected with shRNA‑IGF2BP2 exhibited enhanced cell viability, reduced IL‑1β and IL‑18 concentrations, decreased GSDMD activation in addition to reduced expression levels of caspase 4 and cleaved‑caspase 1 following stimulation with 1 µg/ml LPS. Concomitantly, the effects of IGF2BP2 silencing on caspase 4 expression were higher compared with those noted on caspase 1. In addition, binding of IGF2BP2 to caspase 4 RNA was also observed. To conclude, data from the present study suggest that IGF2BP2 knockdown inhibited LPS‑induced Beas‑2B cell inflammation by targeting caspase 4, thereby inhibiting the non‑canonical pyroptosis pathway.

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1	Perner A, Gordon AC, De Backer D, Dimopoulos G, Russell JA, Lipman J, Jensen JU, Myburgh J, Singer M, Bellomo R and Walsh T: Sepsis: Frontiers in diagnosis, resuscitation and antibiotic therapy. Intensive Care Med. 42:1958–1969. 2016.PubMed/NCBI View Article : Google Scholar
2	Mehaffey JH, Charles EJ, Schubert S, Salmon M, Sharma AK, Money D, Stoler MH, Laubach VE, Tribble CG, Roeser ME and Kron IL: In vivo lung perfusion rehabilitates sepsis-induced lung injury. J Thorac Cardiovasc Surg. 155:440–448 e442. 2018.PubMed/NCBI View Article : Google Scholar
3	Aji G, Li F, Chen J, Leng F, Hu K, Cheng Z, Luo Y, Xu X, Zhang J and Lu Z: Upregulation of PCP4 in human aldosterone-producing adenomas fosters human adrenocortical tumor cell growth via AKT and AMPK pathway. Int J Clin Exp Pathol. 11:1197–1207. 2018.PubMed/NCBI
4	Van Opdenbosch N and Lamkanfi M: Caspases in cell death, inflammation, and disease. Immunity. 50:1352–1364. 2019.PubMed/NCBI View Article : Google Scholar
5	Robinson N, Ganesan R, Hegedűs C, Kovács K, Kufer TA and Virág L: Programmed necrotic cell death of macrophages: Focus on pyroptosis, necroptosis, and parthanatos. Redox Biol. 26(101239)2019.PubMed/NCBI View Article : Google Scholar
6	Hersoug LG, Møller P and Loft S: Role of microbiota-derived lipopolysaccharide in adipose tissue inflammation, adipocyte size and pyroptosis during obesity. Nutr Res Rev. 31:153–163. 2018.PubMed/NCBI View Article : Google Scholar
7	Huang X, Zhang H, Guo X, Zhu Z, Cai H and Kong X: Insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) in cancer. J Hematol Oncol. 11(88)2018.PubMed/NCBI View Article : Google Scholar
8	Wang Y, Lu JH, Wu QN, Jin Y, Wang DS, Chen YX, Liu J, Luo XJ, Meng Q, Pu HY, et al: lncRNA LINRIS stabilizes IGF2BP2 and promotes the aerobic glycolysis in colorectal cancer. Mol Cancer. 18(174)2019.PubMed/NCBI View Article : Google Scholar
9	Li T, Hu PS, Zuo Z, Lin JF, Li X, Wu QN, Chen ZH, Zeng ZL, Wang F, Zheng J, et al: METTL3 facilitates tumor progression via an m(6)A-IGF2BP2-dependent mechanism in colorectal carcinoma. Mol Cancer. 18(112)2019.PubMed/NCBI View Article : Google Scholar
10	Dai N: The diverse functions of IMP2/IGF2BP2 in metabolism. Trends Endocrinol Metab. 31:670–679. 2020.PubMed/NCBI View Article : Google Scholar
11	Simon Y, Kessler SM, Bohle RM, Haybaeck J and Kiemer AK: The insulin-like growth factor 2 (IGF2) mRNA-binding protein p62/IGF2BP2-2 as a promoter of NAFLD and HCC? Gut. 63:861–863. 2014.PubMed/NCBI View Article : Google Scholar
12	Baker PJ, Boucher D, Bierschenk D, Tebartz C, Whitney PG, D'Silva DB, Tanzer MC, Monteleone M, Robertson AAB, Cooper MA, et al: NLRP3 inflammasome activation downstream of cytoplasmic LPS recognition by both caspase-4 and caspase-5. Eur J Immunol. 45:2918–2926. 2015.PubMed/NCBI View Article : Google Scholar
13	Gonzalez-Juarbe N, Bradley KM, Riegler AN, Reyes LF, Brissac T, Park SS, Restrepo MI and Orihuela CJ: Bacterial pore-forming toxins promote the activation of caspases in parallel to necroptosis to enhance alarmin release and inflammation during pneumonia. Sci Rep. 8(5846)2018.PubMed/NCBI View Article : Google Scholar
14	Zaslona Z, Flis E, Wilk MM, Carroll RG, Palsson-McDermott EM, Hughes MM, Diskin C, Banahan K, Ryan DG, Hooftman A, et al: Caspase-11 promotes allergic airway inflammation. Nat Commun. 11(1055)2020.PubMed/NCBI View Article : Google Scholar
15	Srisaowakarn C, Pudla M, Ponpuak M and Utaisincharoen P: Caspase-4 mediates restriction of burkholderia pseudomallei in human alveolar epithelial cells. Infect Immun. 88(e00868)2020.PubMed/NCBI View Article : Google Scholar
16	Bao Z, Chen L and Guo S: Knockdown of SLC34A2 inhibits cell proliferation, metastasis, and elevates chemosensitivity in glioma. J Cell Biochem. 120:10205–10214. 2019.PubMed/NCBI View Article : Google Scholar
17	Ko IG, Hwang JJ, Chang BS, Kim HS, Jin JJ, Hwang L, Kim CJ and Choi CW: Polydeoxyribonucleotide ameliorates lipopolysaccharide-induced acute lung injury via modulation of the MAPK/NF-kB signaling pathway in rats. Int Immunopharmacol. 83(106444)2020.PubMed/NCBI View Article : Google Scholar
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.PubMed/NCBI View Article : Google Scholar
19	Dou YQ, Kong P, Li CL, Sun HX, Li WW, Yu Y, Nie L, Zhao LL, Miao SB, Li XK, et al: Smooth muscle SIRT1 reprograms endothelial cells to suppress angiogenesis after ischemia. Theranostics. 10:1197–1212. 2020.PubMed/NCBI View Article : Google Scholar
20	Lv X, Zhou X, Yan J, Jiang J and Jiang H: Propofol inhibits LPS-induced apoptosis in lung epithelial cell line, BEAS-2B. Biomed Pharmacother. 87:180–187. 2017.PubMed/NCBI View Article : Google Scholar
21	Aaltomaa S, Kärjä V, Lipponen P, Isotalo T, Kankkunen JP, Talja M and Mokka R: Reduced alpha- and beta-catenin expression predicts shortened survival in local prostate cancer. Anticancer Res. 25:4707–4712. 2005.PubMed/NCBI
22	He C, Zhao Y, Jiang X, Liang X, Yin L, Yin Z, Geng Y, Zhong Z, Song X, Zou Y, et al: Protective effect of Ketone musk on LPS/ATP-induced pyroptosis in J774A.1 cells through suppressing NLRP3/GSDMD pathway. Int Immunopharmacol. 71:328–335. 2019.PubMed/NCBI View Article : Google Scholar
23	Wan BS, Cheng M and Zhang L: Insulin-like growth factor 2 mRNA-binding protein 1 promotes cell proliferation activation of AKT and is directly targeted by microRNA-494 in pancreatic cancer. World J Gastroenterol. 25:6063–6076. 2019.PubMed/NCBI View Article : Google Scholar
24	Abrenica B, AlShaaban M and Czubryt MP: The A-kinase anchor protein AKAP121 is a negative regulator of cardiomyocyte hypertrophy. J Mol Cell Cardiol. 46:674–681. 2009.PubMed/NCBI View Article : Google Scholar
25	McMullen ER, Gonzalez ME, Skala SL, Tran M, Thomas D, Djomehri SI, Burman B, Kidwell KM and Kleer CG: CCN6 regulates IGF2BP2 and HMGA2 signaling in metaplastic carcinomas of the breast. Breast Cancer Res Treat. 172:577–586. 2018.PubMed/NCBI View Article : Google Scholar
26	Guiot J, Henket M, Corhay JL, Moermans C and Louis R: Sputum biomarkers in IPF: Evidence for raised gene expression and protein level of IGFBP-2, IL-8 and MMP-7. PLoS One. 12(e0171344)2017.PubMed/NCBI View Article : Google Scholar
27	Liu Y, Zhang B, Cao WB, Wang HY, Niu L and Zhang GZ: Study on clinical significance of lncRNA EGOT expression in colon cancer and its effect on autophagy of colon cancer cells. Cancer Manag Res. 12:13501–13512. 2020.PubMed/NCBI View Article : Google Scholar
28	Fei S, Cao L and Pan L: microRNA-3941 targets IGF2 to control LPS-induced acute pneumonia in A549 cells. Mol Med Rep. 17:4019–4026. 2018.PubMed/NCBI View Article : Google Scholar