miR-450b-5p Regulates Adhesion and Migration of Human Gingival Epithelial Cells Induced by IL-1β

doi:10.13701/j.cnki.kqyxyj.2022.06.016

Abstract

Abstract: Objective: To investigate the effect of miR-450b-5p on the adhesion and migration of human gingival epithelial cells (HGEs) regulated by inflammatory factor IL-1β. Methods: HGEs were cultured in vitro and starved for 12 hours in logarithmic growth period, stimulated by 5 ng/mL IL-1β for 3 hours. Total RNA was extracted by RNAiso Plus and analyzed by small RNA sequencing and qRT-PCR. The migration of HGEs induced by IL-1β was observed by cell scratch assay. After transfected with miR-450b-5p mimic, inhibitor, and NC lentivirus plasmid, the expressions of miR-450b-5p, Integrinβ6 (ITGβ6), Lamininγ2 (LAMC2), and matrix metalloproteinase-9 (MMP-9) in HGEs regulated by IL-1β were detected by qRT-PCR. Results: Sequencing results and qRT-PCR showed that IL-1β decreased the expression of miR-450b-5p. The expression of ITGβ6, LAMC2, and MMP-9 were increased after IL-1β induction. The expressions of ITGβ6, LAMC2, and MMP-9 genes were inhibited and the increased effect induced by IL-1β was partially reversed by overexpression of miR-450b-5p. However, the opposite result was obtained by knockdown of miR-450b-5p. Cell scratch assay showed that migration of HGEs induced by IL-1β was inhibited by overexpression of miR-450b-5p. Conclusion: MiR-450b-5p inhibits HGEs adhesion and migration induced by IL-1β, suggesting that miR-450b-5p plays an important role in maintaining HGEs stability, inhibiting cell migration and protecting the gingival epithelial barrier.

Key words: IL-1β, miR-450b-5p, lamininγ2, ITGβ6, MMP-9

YU Cuicui, YE Xiaoyuan, DONG Zhiheng, WANG Aiqin, GAO Yan, GAO Yuguang. miR-450b-5p Regulates Adhesion and Migration of Human Gingival Epithelial Cells Induced by IL-1β[J]. Journal of Oral Science Research, 2022, 38(6): 572-577.

References

[1] Da SF, Vasconcelos A, de Carvalho FL, et al. Association between the rs1143634 polymorphism in interleukin-1B and chronic periodontitis: Results from a meta-analysis composed by 54 case/control studies [J]. Gene, 2018, 668:97-106.
[2] Groeger SE, Meyle J. Epithelial barrier and oral bacterial infection[J]. Periodontol 2000, 2015, 69(1):46-67.
[3] Wang SS, Tang YL, Pang X, et al. The maintenance of an oral epithelial barrier [J]. Life Sci, 2019, 227:129-136.
[4] Fields JK, Gunther S, Sundberg EJ. Structural basis of IL-1 family cytokine signaling [J]. Front Immunol, 2019, 10:1412.
[5] Stephenson J, Nutma E, van der Valk P, et al. Inflammation in CNS neurodegenerative diseases [J]. Immunology, 2018, 154(2):204-219.
[6] Ayazi G, Pirayesh M, Yari K. Analysis of interleukin-1beta gene polymorphism and its association with generalized aggressive periodontitis disease [J]. DNA Cell Biol, 2013, 32(7):409-413.
[7] Chen SC, Constantinides C, Kebschull M, et al. MicroRNAs regulate cytokine responses in gingival epithelial cells [J]. Infect Immun, 2016, 84(12):3282-3289.
[8] How KY, Song KP, Chan KG. Porphyromonas gingivalis: An overview of periodontopathic pathogen below the gum Line [J]. Front Microbiol, 2016, 7:53.
[9] Kato H, Taguchi Y, Tominaga K, et al. Porphyromonas gingivalis LPS inhibits osteoblastic differentiation and promotes pro-inflammatory cytokine production in human periodontal ligament stem cells [J]. Arch Oral Biol, 2014, 59(2):167-175.
[10] Akkaoui J, Yamada C, Duarte C, et al. Contribution of porphyromonas gingivalis lipopolysaccharide to experimental periodontitis in relation to aging [J]. Geroscience, 2021, 43(1):367-376.
[11] Petrasek J, Bala S, Csak T, et al. IL-1 receptor antagonist ameliorates inflammasome-dependent alcoholic steatohepatitis in mice [J]. J Clin Invest, 2012, 122(10):3476-3489.
[12] Boraschi D, Tagliabue A. The interleukin-1 receptor family [J]. Semin Immunol, 2013, 25(6):394-407.
[13] Li H, Shen S, Chen X, et al. miR-450b-5p loss mediated KIF26B activation promoted hepatocellular carcinoma progression by activating PI3K/AKT pathway [J]. Cancer Cell Int, 2019, 19:205.
[14] Chang YC, Sabourin CL, Lu SE, et al. Upregulation of gamma-2 laminin-332 in the mouse ear vesicant wound model [J]. J Biochem Mol Toxicol, 2009, 23(3):172-184.
[15] Zboralski D, Warscheid B, Klein-Scory S, et al. Uncoupled responses of Smad4-deficient cancer cells to TNFalpha result in secretion of monomeric laminin-gamma2 [J]. Mol Cancer, 2010, 9(1):65.
[16] Koivisto L, Bi J, Hakkinen L, et al. Integrin alphavbeta6: Structure, function and role in health and disease [J]. Int J Biochem Cell Biol, 2018, 99:186-196.
[17] Niu J, Li Z. The roles of integrin alphavbeta6 in cancer [J]. Cancer Lett, 2017, 403:128-137.
[18] Elkington PT, O'Kane CM, Friedland JS. The paradox of matrix metalloproteinases in infectious disease [J]. Clin Exp Immunol, 2005, 142(1):12-20.
[19] Zhang H, Liu L, Jiang C, et al. MMP-9 protects against LPS-induced inflammation in osteoblasts [J]. Innate Immun, 2020, 26(4):259-269.