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Inhibition of Stat3 signaling pathway decreases TNF-α-induced autophagy in cementoblasts

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Abstract

Autophagy is a self-digestive process that eliminates impaired or aged proteins and potentially toxic intracellular components to maintain homeostasis. We previously demonstrated that TNF-α played a critical role in cementoblast differentiation, mineralization and apoptosis; however, the effect of TNF-α on cementoblast autophagy has remained unclear. In this study, an elevated immunofluorescence signal of LC3B and autophagic vacuoles, autophagosomes and autolysosomes were detected under TNF-α stimulation in OCCM-30 cells. Autophagy-related genes and proteins, Beclin-1, LC3A and Atg-5, were significantly upregulated by TNF-α in a time- and concentration-dependent manner. During this process, the activity of Stat3 was dramatically enhanced and when the activity of Stat3 was blocked by either a specific chemical inhibitor or siRNA transfection before TNF-α stimulation, the TNF-α-induced upregulation of autophagy-related genes and proteins was strongly inhibited. Our results suggest that TNF-α induced autophagy in cementoblasts was dependent, or partially dependent on the activity of Stat3 signaling pathway.

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References

  • An Y, Liu W, Xue P, Zhang Y, Wang Q, Jin Y (2016) Increased autophagy is required to protect periodontal ligament stem cells from apoptosis in inflammatory microenvironment. J Clin Periodontol 43:618–625

    Article  Google Scholar 

  • Bartolome A, Lopez-Herradon A, Portal-Nunez S, Garcia-Aguilar A, Esbrit P, Benito M, Guillen C (2013) Autophagy impairment aggravates the inhibitory effects of high glucose on osteoblast viability and function. Biochem J 455:329–337

    Article  CAS  Google Scholar 

  • Beertsen W, Mcculloch CA, Sodek J (1997) The periodontal ligament: a unique, multifunctional connective tissue. Periodontol 2000 13:20–40

    Article  CAS  Google Scholar 

  • Brenner C, Galluzzi L, Kepp O, Kroemer G (2013) Decoding cell death signals in liver inflammation. J Hepatol 59:583–594

    Article  CAS  Google Scholar 

  • Bromberg J (2002) Stat proteins and oncogenesis. J Clin Investig 109:1139–1142

    Article  CAS  Google Scholar 

  • Chen XT, Ying Z, Lin X, Lin HX, Wu JH, Li MF, Song LB (2013) Acylglycerol kinase augments jak2/stat3 signaling in esophageal squamous cells. J Clin Investig 123:2576–2589

    Article  CAS  Google Scholar 

  • D’errico JA, Berry JE, Ouyang H, Strayhorn CL, Windle JJ, Somerman MJ (2000) Employing a transgenic animal model to obtain cementoblasts in vitro. J Periodontol 71:63–72

    Article  Google Scholar 

  • Dang S, Xu H, Xu C, Cai W, Li Q, Cheng Y, Jin M, Wang RX, Peng Y, Zhang Y, Wu C, He X, Wan B, Zhang Y (2014) Autophagy regulates the therapeutic potential of mesenchymal stem cells in experimental autoimmune encephalomyelitis. Autophagy 10:1301–1315

    Article  Google Scholar 

  • Dang S, Yu ZM, Zhang CY, Zheng J, Li KL, Wu Y, Qian LL, Yang ZY, Li XR, Zhang Y (2015) Autophagy promotes apoptosis of mesenchymal stem cells under inflammatory microenvironment. Stem Cell Res Ther 6:247–253

    Article  Google Scholar 

  • Diekwisch TG (2001) The developmental biology of cementum. Int J Dev Biol 45:695–706

    CAS  PubMed  Google Scholar 

  • Djavaheri-Mergny M, Amelotti M, Mathieu J, Besancon F, Bauvy C, Souquere S, Pierron G, Codogno P (2006) Nf-kappab activation represses tumor necrosis factor-alpha-induced autophagy. J Biol Chem 281:30373–30382

    Article  CAS  Google Scholar 

  • Du TA (2012) Autophagy: Stat3 maintains order. Nat Rev Mol Cell Biol 13:754–760

    Google Scholar 

  • Ertugrul AS, Sahin H, Dikilitas A, Alpaslan N, Bozoglan A (2013) Comparison of ccl28, interleukin-8, interleukin-1beta and tumor necrosis factor-alpha in subjects with gingivitis, chronic periodontitis and generalized aggressive periodontitis. J Periodontal Res 48:44–51

    Article  CAS  Google Scholar 

  • Fan TF, Wu TF, Bu LL, Ma SR, Li YC, Mao L, Sun ZJ, Zhang WF (2016) Dihydromyricetin promotes autophagy and apoptosis through ros-stat3 signaling in head and neck squamous cell carcinoma. Oncotarget 7:59691–59703

    PubMed  PubMed Central  Google Scholar 

  • Feng Y, Ke C, Tang Q, Dong H, Zheng X, Lin W, Ke J, Huang J, Yeung SC, Zhang H (2014) Metformin promotes autophagy and apoptosis in esophageal squamous cell carcinoma by downregulating stat3 signaling. Cell Death Dis 5:e1088–e1099

    Article  CAS  Google Scholar 

  • Gao B, Wang H, Lafdil F, Feng D (2012) Stat proteins—key regulators of anti-viral responses, inflammation, and tumorigenesis in the liver. J Hepatol 57:430–441

    Article  CAS  Google Scholar 

  • Ha SW, Weitzmann MN, Beck GR Jr (2014) Bioactive silica nanoparticles promote osteoblast differentiation through stimulation of autophagy and direct association with lc3 and p62. ACS Nano 8:5898–5910

    Article  CAS  Google Scholar 

  • He M, Wang C, Sun JH, Liu Y, Wang H, Zhao JS, Li YF, Chang H, Hou JM, Song JN (2017) Roscovitine attenuates intimal hyperplasia via inhibiting nf-κb and stat3 activation induced by tnf-α in vascular smooth muscle cells. Biochem Pharmacol 137:51–60

    Article  CAS  Google Scholar 

  • Jude B, Vetel S, Girouxmetges MA, Pennec JP (2017) Rapid negative inotropic effect induced by tnf-α in rat heart perfused related to pkc activation. Cytokine 107:65–69

    Article  Google Scholar 

  • Kang R, Tang D, Lotze MT, Iii HJZ (2012) Ager/rage-mediated autophagy promotes pancreatic tumorigenesis and bioenergetics through the il6-pstat3 pathway. Autophagy 8:989–991

    Article  CAS  Google Scholar 

  • Kim IR, Kim SE, Baek HS, Kim BJ, Kim CH, Chung IK, Park BS, Shin SH (2016) The role of kaempferol-induced autophagy on differentiation and mineralization of osteoblastic mc3t3-e1 cells. BMC Complement Altern Med 16:333–342

    Article  Google Scholar 

  • Kim J, Lee HW, Rhee DK, Paton JC, Pyo S (2017) Pneumolysin-induced autophagy contributes to inhibition of osteoblast differentiation through downregulation of sp1 in human osteosarcoma cells. Biochim Biophys Acta 1861:2663–2673

    Article  CAS  Google Scholar 

  • Lee BH, Hsu WH, Liao TH, Pan TM (2011) The monascus metabolite monascin against tnf-alpha-induced insulin resistance via suppressing ppar-gamma phosphorylation in c2c12 myotubes. Food Chem Toxicol 49:2609–2617

    Article  CAS  Google Scholar 

  • Levine B, Klionsky DJ (2004) Development by self-digestion: Molecular mechanisms and biological functions of autophagy. Dev Cell 6:463–477

    Article  CAS  Google Scholar 

  • Levine B, Kroemer G (2008) Autophagy in the pathogenesis of disease. Cell 132:27–42

    Article  CAS  Google Scholar 

  • Levine B, Mizushima N, Virgin HW (2011) Autophagy in immunity and inflammation. Nature 469:323–335

    Article  CAS  Google Scholar 

  • Levy DE, Lee CK (2002) What does stat3 do? J Clin Investig 109:1143–1148

    Article  CAS  Google Scholar 

  • Liu F, Fang F, Yuan H, Yang D, Chen Y, Williams L, Goldstein SA, Krebsbach PH, Guan JL (2013) Suppression of autophagy by fip200 deletion leads to osteopenia in mice through the inhibition of osteoblast terminal differentiation. J Bone Miner Res 28:2414–2430

    Article  CAS  Google Scholar 

  • Mizushima N, Noda T, Yoshimori T, Tanaka Y, Ishii T, George MD, Klionsky DJ, Ohsumi M, Ohsumi Y (1998) A protein conjugation system essential for autophagy. Nature 395:395–398

    Article  CAS  Google Scholar 

  • Mizushima N, Yoshimori T, Levine B (2010) Methods in mammalian autophagy research. Cell 140:313–326

    Article  CAS  Google Scholar 

  • Mori T, Miyamoto T, Yoshida H, Asakawa M, Kawasumi M, Kobayashi T, Morioka H, Chiba K, Toyama Y, Yoshimura A (2011) Il-1beta and tnfalpha-initiated il-6-stat3 pathway is critical in mediating inflammatory cytokines and rankl expression in inflammatory arthritis. Int Immunol 23:701–712

    Article  CAS  Google Scholar 

  • Oates TW, Graves DT, Cochran DL (2002) Clinical, radiographic and biochemical assessment of il-1/tnf-alpha antagonist inhibition of bone loss in experimental periodontitis. J Clin Periodontol 29:137–143

    Article  CAS  Google Scholar 

  • Oh SY, Choi SJ, Kim KH, Cho EY, Kim JH, Roh CR (2008) Autophagy-related proteins, lc3 and beclin-1, in placentas from pregnancies complicated by preeclampsia. Reprod Sci 15:912–920

    Article  CAS  Google Scholar 

  • Oka H, Miyauchi M, Sakamoto K, Moriwaki S, Niida S, Noguchi K, Somerman MJ, Takata T (2007) Pge2 activates cementoclastogenesis by cementoblasts via ep4. J Dent Res 86:974–979

    Article  CAS  Google Scholar 

  • Padial-Molina M, Volk SL, Rodriguez JC, Marchesan JT, Galindo-Moreno P, Rios HF (2013) Tumor necrosis factor-alpha and porphyromonas gingivalis lipopolysaccharides decrease periostin in human periodontal ligament fibroblasts. J Periodontol 84:694–703

    Article  CAS  Google Scholar 

  • Pei F, Lin H, Liu H, Li L, Zhang L, Chen Z (2015) Dual role of autophagy in lipopolysaccharide-induced preodontoblastic cells. J Dent Res 94:175–182

    Article  CAS  Google Scholar 

  • Pierrefite-Carle V, Santucci-Darmanin S, Breuil V, Camuzard O, Carle GF (2015) Autophagy in bone: self-eating to stay in balance. Ageing Res Rev 24:206–217

    Article  Google Scholar 

  • Qin B, Zhuo Z, He J, Yan C, Ding S (2015) Il-6 inhibits starvation-induced autophagy via the stat3/bcl-2 signaling pathway. Sci Rep 5:15701–15710

    Article  CAS  Google Scholar 

  • Sandoval H, Thiagarajan P, Dasgupta SK, Schumacher A, Prchal JT, Chen M, Wang J (2008) Essential role for nix in autophagic maturation of erythroid cells. Nature 454:232–235

    Article  CAS  Google Scholar 

  • Shapiro IM, Layfield R, Lotz M, Settembre C, Whitehouse C (2014) Boning up on autophagy: the role of autophagy in skeletal biology. Autophagy 10:7–19

    Article  CAS  Google Scholar 

  • Shen S, Niso-Santano M, Adjemian S, Takehara T, Malik SA, Minoux H, Souquere S, Marino G, Lachkar S, Senovilla L, Galluzzi L, Kepp O, Pierron G, Maiuri MC, Hikita H, Kroemer R, Kroemer G (2012) Cytoplasmic stat3 represses autophagy by inhibiting pkr activity. Mol Cell 48:667–680

    Article  CAS  Google Scholar 

  • Shen S, Niso-Santano M, Adjemian S, Takehara T, Malik SA, Minoux H, Souquere S, Mariño G, Lachkar S, Senovilla L (2013) Cytoplasmic stat3 represses autophagy by inhibiting pkr activity. Mol Cell 48:667–680

    Article  Google Scholar 

  • Takano-Ohmuro H, Mukaida M, Kominami E, Morioka K (2000) Autophagy in embryonic erythroid cells: its role in maturation. Eur J Cell Biol 79:759–764

    Article  CAS  Google Scholar 

  • Wang YL, He H, Liu ZJ, Cao ZG, Wang XY, Yang K, Fang Y, Han M, Zhang C, Huo FY (2015) Effects of tnf-alpha on cementoblast differentiation, mineralization, and apoptosis. J Dent Res 94:1225–1232

    Article  CAS  Google Scholar 

  • Wong PF, Jamal J, Tong KL, Khor ES, Yeap CE, Jong HL, Lee ST, Mustafa MR, Abubakar S (2017) Deregulation of hsa-mir-20b expression in tnf-alpha-induced premature senescence of human pulmonary microvascular endothelial cells. Microvasc Res 114:26–33

    Article  CAS  Google Scholar 

  • Wu J, Guo J, Cao Q, Wang Y, Chen J, Wang Z, Yuan Z (2017) Autophagy impacts on oxaliplatin-induced hepatocarcinoma apoptosis via the il-17/il-17r-jak2/stat3 signaling pathway. Oncol Lett 13:770–776

    Article  CAS  Google Scholar 

  • Ye YC, Wang HJ, Yu L, Tashiro S, Onodera S, Ikejima T (2012) Rip1-mediated mitochondrial dysfunction and ros production contributed to tumor necrosis factor alpha-induced l929 cell necroptosis and autophagy. Int Immunopharmacol 14:674–682

    Article  CAS  Google Scholar 

  • Yoon S, Woo SU, Kang JH, Kim K, Kwon MH, Park S, Shin HJ, Gwak HS, Chwae YJ (2010) Stat3 transcriptional factor activated by reactive oxygen species induces il6 in starvation-induced autophagy of cancer cells. Autophagy 6:1125–1138

    Article  CAS  Google Scholar 

  • You L, Wang Z, Li H, Shou J, Jing Z, Xie J, Sui X, Pan H, Han W (2015) The role of stat3 in autophagy. Autophagy 11:729–739

    Article  CAS  Google Scholar 

  • Yu H, Jove R (2004) The stats of cancer—new molecular targets come of age. Nat Rev Cancer 4:97–105

    Article  CAS  Google Scholar 

  • Yu H, Pardoll D, Jove R (2009) Stats in cancer inflammation and immunity: a leading role for stat3. Nat Rev Cancer 9:798–809

    Article  CAS  Google Scholar 

  • Zhao D, Yuan H, Yi F, Meng C, Zhu Q (2014) Autophagy prevents doxorubicininduced apoptosis in osteosarcoma. Mol Med Rep 9:1975–1981

    Article  CAS  Google Scholar 

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Funding

This study was financially supported by grants from the National Natural Science Foundation of China (Nos. 81671020 and 81200811) and the Natural Science Foundation of Hubei Province (No. 2015CFB404)

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Authors and Affiliations

  1. Department of Orthodontics, The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Luoyu Road #237, Hongshan District, Wuhan, 430079, China

    Leilei Wang, Yunlong Wang, Mingyuan Du, Zhijian Liu, Zhengguo Cao, Yunru Hao & Hong He

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  1. Leilei Wang
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  2. Yunlong Wang
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  3. Mingyuan Du
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  4. Zhijian Liu
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  5. Zhengguo Cao
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  6. Yunru Hao
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  7. Hong He
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Contributions

Study design: L.L. Wang, Y.L. Wang, H. He, Z.J. Liu, Z.G. Cao and M.Y. Du. Data Collection and analysis: L.L. Wang, and Y.R. Hao. Contribution of new reagents or analytical tools: M.Y. Du. Manuscript preparation: L.L. Wang, Y.L. Wang and H. He.

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Correspondence to Hong He.

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Wang, L., Wang, Y., Du, M. et al. Inhibition of Stat3 signaling pathway decreases TNF-α-induced autophagy in cementoblasts. Cell Tissue Res 374, 567–575 (2018). https://doi.org/10.1007/s00441-018-2890-2

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