Skip to main content

Advertisement

SpringerLink
Log in

Phosphorylation and nuclear translocation of integrin β4 induced by a chemical small molecule contribute to apoptosis in vascular endothelial cells

  • Original Paper
  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Integrin β4 and its Y-1494 phosphorylation play an important role in cell signaling. We found a small molecule, ethyl1-(3-(4-chlorophenoxy)-2-hydroxypropyl)-3-(4-chlorophenyl)-1H-pyrazole-5-carboxylate (ECPC), that could elevate the levels of KIT ligand (KITLG), interleukin 8 (IL-8), prostaglandin-endoperoxide synthase 2 (PTGS2) and activating transcription factor 3 (ATF3) and promote apoptosis in vascular endothelial cells (VECs) through integrin β4. We investigated the underlying mechanism of integrin β4 participating in this process. ECPC treatment increased the phosphorylation of Y-1494 in the integrin β4 cytoplasmic domain via a well-known receptor tyrosine kinase, fibroblast growth factor receptor 1 (FGFR1), and integrin β4 translocated from the cytoplasm to nucleus. With suppression of Y-1494 phosphorylation by FGF-2 or siRNA of FGFR1, ECPC failed to promote integrin β4 nuclear translocation and could not increase the expression of KITLG, IL-8, PTGS2 or ATF3. Y-1494 phosphorylation and nuclear translocation of integrin β4 may be important during ECPC-induced apoptosis in VECs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

ECPC:

Ethyl1-(3-(4-chlorophenoxy)-2-hydroxypropyl)-3-(4-chlorophenyl)-1H-pyrazole-5-carboxylate

VEC:

Vascular endothelial cell

HUVECs:

Human umbilical vein endothelial cells

IL-8:

Interleukin 8

RNAi:

RNA interference

siRNA:

Small interfering RNA

FBS:

Fetal bovine serum

FGF-2:

Fibroblast growth factor-2

FGFR1:

Fibroblast growth factor receptor 1

RTK:

Receptor tyrosine kinase

PTGS2:

Prostaglandin-endoperoxide synthase 2

ATF3:

Activating transcription factor 3

EGFR:

Epidermal growth factor receptor

DMSO:

Dimethyl sulfoxide

References

  1. de Pereda JM, Ortega E, Alonso-Garcia N, Gomez-Hernandez M, Sonnenberg A (2009) Advances and perspectives of the architecture of hemidesmosomes: lessons from structural biology. Cell Adh Migr 3:361–364

    Article  PubMed  Google Scholar 

  2. Wang L, Dong Z, Zhang Y, Miao J (2012) The roles of integrin beta4 in vascular endothelial cells. J Cell Physiol 227:474–478

    Article  PubMed  CAS  Google Scholar 

  3. Borradori L, Sonnenberg A (1999) Structure and function of hemidesmosomes: more than simple adhesion complexes. J Invest Dermatol 112:411–418

    Article  PubMed  CAS  Google Scholar 

  4. Lipscomb EA, Mercurio AM (2005) Mobilization and activation of a signaling competent alpha6beta4integrin underlies its contribution to carcinoma progression. Cancer Metastasis Rev 24:413–423

    Article  PubMed  CAS  Google Scholar 

  5. Margadant C, Frijns E, Wilhelmsen K, Sonnenberg A (2008) Regulation of hemidesmosome disassembly by growth factor receptors. Curr Opin Cell Biol 20:589–596

    Article  PubMed  CAS  Google Scholar 

  6. Shaw LM (2001) Identification of insulin receptor substrate 1 (IRS-1) and IRS-2 as signaling intermediates in the alpha6beta4 integrin-dependent activation of phosphoinositide 3-OH kinase and promotion of invasion. Mol Cell Biol 21:5082–5093

    Article  PubMed  CAS  Google Scholar 

  7. Choy JC, Granville DJ, Hunt DW, McManus BM (2001) Endothelial cell apoptosis: biochemical characteristics and potential implications for atherosclerosis. J Mol Cell Cardiol 33:1673–1690

    Article  PubMed  CAS  Google Scholar 

  8. Miao JY, Araki S, Kaji K, Hayashi H (1997) Integrin beta4 is involved in apoptotic signal transduction in endothelial cells. Biochem Biophys Res Commun 233:182–186

    Article  PubMed  CAS  Google Scholar 

  9. Zhao Q, Araki S, Zhang S, Miao J (2004) Rattlesnake venom induces apoptosis by stimulating PC-PLC and upregulating the expression of integrin beta4, P53 in vascular endothelial cells. Toxicon 44:161–168

    Article  PubMed  CAS  Google Scholar 

  10. Zhao J, Miao J, Zhao B, Zhang S (2005) Upregulating of Fas, integrin beta4 and P53 and depressing of PC-PLC activity and ROS level in VEC apoptosis by safrole oxide. FEBS Lett 579:5809–5813

    Article  PubMed  CAS  Google Scholar 

  11. Fan C, Su H, Zhao J, Zhao B, Zhang S, Miao J (2010) A novel copper complex of salicylaldehyde pyrazole hydrazone induces apoptosis through up-regulating integrin beta4 in H322 lung carcinoma cells. Eur J Med Chem 45:1438–1446

    Article  PubMed  CAS  Google Scholar 

  12. Zhang L, Zhu X, Zhao B, Zhao J, Zhang Y, Zhang S et al (2008) A novel isochroman derivative inhibited apoptosis in vascular endothelial cells through depressing the levels of integrin beta4, p53 and ROS. Vascul Pharmacol 48:63–69

    Article  PubMed  Google Scholar 

  13. Dong Z, Wang L, Xu J, Li Y, Zhang Y, Zhang S et al (2009) Promotion of autophagy and inhibition of apoptosis by low concentrations of cadmium in vascular endothelial cells. Toxicol In Vitro 23:105–110

    Article  PubMed  CAS  Google Scholar 

  14. Wei F, Zhao BX, Huang B, Zhang L, Sun CH, Dong WL et al (2006) Design, synthesis, and preliminary biological evaluation of novel ethyl 1-(2′-hydroxy-3′-aroxypropyl)-3-aryl-1H-pyrazole-5-carboxylate. Bioorg Med Chem Lett 16:6342–6347

    Article  PubMed  CAS  Google Scholar 

  15. Jaffe EA, Nachman RL, Becker CG, Minick CR (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52:2745–2756

    Article  PubMed  CAS  Google Scholar 

  16. Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493–501

    Article  PubMed  CAS  Google Scholar 

  17. Wang N, Sun C, Huo S, Zhang Y, Zhao J, Zhang S et al (2008) Cooperation of phosphatidylcholine-specific phospholipase C and basic fibroblast growth factor in the neural differentiation of mesenchymal stem cells in vitro. Int J Biochem Cell Biol 40:294–306

    Article  PubMed  CAS  Google Scholar 

  18. Patterson TA, Lobenhofer EK, Fulmer-Smentek SB, Collins PJ, Chu TM, Bao W et al (2006) Performance comparison of one-color and two-color platforms within the MicroArray Quality Control (MAQC) project. Nat Biotechnol 24:1140–1150

    Article  PubMed  CAS  Google Scholar 

  19. Wang YN, Yamaguchi H, Hsu JM, Hung MC (2010) Nuclear trafficking of the epidermal growth factor receptor family membrane proteins. Oncogene 29:3997–4006

    Article  PubMed  CAS  Google Scholar 

  20. Du AY, Zhao BX, Yin DL, Zhang SL, Miao JY (2005) Discovery of a novel small molecule, 1-ethoxy-3-(3,4-methylenedioxyphenyl)-2-propanol, that induces apoptosis in A549 human lung cancer cells. Bioorg Med Chem 13:4176–4183

    Article  PubMed  CAS  Google Scholar 

  21. Sun C, Liu X, Qi L, Xu J, Zhao J, Zhang Y et al (2010) Modulation of vascular endothelial cell senescence by integrin beta4. J Cell Physiol 225:673–681

    Article  PubMed  CAS  Google Scholar 

  22. Ge D, Jing Q, Meng N, Su L, Zhang Y, Zhang S et al (2011) Regulation of apoptosis and autophagy by sphingosylphosphorylcholine in vascular endothelial cells. J Cell Physiol 226:2827–2833

    Article  PubMed  CAS  Google Scholar 

  23. Wang YN, Lee HH, Lee HJ, Du Y, Yamaguchi H, Hung MC (2012) Membrane-bound trafficking regulates nuclear transport of integral epidermal growth factor receptor (EGFR) and ErbB-2. J Biol Chem 287:16869–16879

    Article  PubMed  CAS  Google Scholar 

  24. Giancotti FG (2007) Targeting integrin beta4 for cancer and anti-angiogenic therapy. Trends Pharmacol Sci 28:506–511

    Article  PubMed  CAS  Google Scholar 

  25. Kazenwadel J, Secker GA, Betterman KL, Harvey NL (2012) In vitro assays using primary embryonic mouse lymphatic endothelial cells uncover key roles for FGFR1 signalling in lymphangiogenesis. PLoS One 7:e40497

    Article  PubMed  CAS  Google Scholar 

  26. Garcia-Maya M, Anderson AA, Kendal CE, Kenny AV, Edwards-Ingram LC, Holladay A et al (2006) Ligand concentration is a driver of divergent signaling and pleiotropic cellular responses to FGF. J Cell Physiol 206:386–393

    Article  PubMed  CAS  Google Scholar 

  27. Huang WC, Chen YJ, Li LY, Wei YL, Hsu SC, Tsai SL et al (2011) Nuclear translocation of epidermal growth factor receptor by Akt-dependent phosphorylation enhances breast cancer-resistant protein expression in gefitinib-resistant cells. J Biol Chem 286:20558–20568

    Article  PubMed  CAS  Google Scholar 

  28. Dutta U, Shaw LM (2008) A key tyrosine (Y1494) in the beta4 integrin regulates multiple signaling pathways important for tumor development and progression. Cancer Res 68:8779–8787

    Article  PubMed  CAS  Google Scholar 

  29. Hung MC, Link W (2011) Protein localization in disease and therapy. J Cell Sci 124:3381–3392

    Article  PubMed  CAS  Google Scholar 

  30. Parkhurst CN, Zampieri N, Chao MV (2010) Nuclear localization of the p75 neurotrophin receptor intracellular domain. J Biol Chem 285:5361–5368

    Article  PubMed  CAS  Google Scholar 

  31. Chen M, Sinha M, Luxon BA, Bresnick AR, O’Connor KL (2009) Integrin alpha6beta4 controls the expression of genes associated with cell motility, invasion, and metastasis, including S100A4/metastasin. J Biol Chem 284:1484–1494

    Article  PubMed  CAS  Google Scholar 

  32. Kligys KR, Wu Y, Hopkinson SB, Kaur S, Platanias LC, Jones JC (2012) α6β4 integrin, a master regulator of expression of integrins in human keratinocytes. J Biol Chem 287:17975–17984

    Article  PubMed  CAS  Google Scholar 

  33. Giri DK, Ali-Seyed M, Li LY, Lee DF, Ling P, Bartholomeusz G et al (2005) Endosomal transport of ErbB-2: mechanism for nuclear entry of the cell surface receptor. Mol Cell Biol 25:11005–11018

    Article  PubMed  CAS  Google Scholar 

  34. Kashyap T, Rabinovitz I (2012) The calcium/calcineurin pathway promotes hemidesmosome stability through inhibition of β4 integrin phosphorylation. J Biol Chem 287:32440–32449

    Article  PubMed  CAS  Google Scholar 

  35. Chioni AM, Grose R (2012) FGFR1 cleavage and nuclear translocation regulates breast cancer cell behavior. J Cell Biol 197:801–817

    Article  PubMed  CAS  Google Scholar 

  36. Stachowiak MK, Fang X, Myers JM, Dunham SM, Berezney R, Maher PA et al (2003) Integrative nuclear FGFR1 signaling (INFS) as a part of a universal “feed-forward-and-gate” signaling module that controls cell growth and differentiation. J Cell Biochem 90:662–691

    Article  PubMed  CAS  Google Scholar 

  37. Beltrami AP, Barlucchi L, Torella D, Baker M, Limana F, Chimenti S et al (2003) Adult cardiac stem cells are multipotent and support myocardial regeneration. Cell 114:763–776

    Article  PubMed  CAS  Google Scholar 

  38. Yan W, Suominen J, Toppari J (2000) Stem cell factor protects germ cells from apoptosis in vitro. J Cell Sci 113(Pt 1):161–168

    PubMed  CAS  Google Scholar 

  39. Weiss N, Deboux C, Chaverot N, Miller F, Baron-Van AE, Couraud PO et al (2010) IL8 and CXCL13 are potent chemokines for the recruitment of human neural precursor cells across brain endothelial cells. J Neuroimmunol 223:131–134

    Article  PubMed  CAS  Google Scholar 

  40. Talavera D, Castillo AM, Dominguez MC, Gutierrez AE, Meza I (2004) IL8 release, tight junction and cytoskeleton dynamic reorganization conducive to permeability increase are induced by dengue virus infection of microvascular endothelial monolayers. J Gen Virol 85:1801–1813

    Article  PubMed  CAS  Google Scholar 

  41. Murray CL, Skelly DT, Cunningham C (2011) Exacerbation of CNS inflammation and neurodegeneration by systemic LPS treatment is independent of circulating IL-1beta and IL-6. J Neuroinflammation 8:50

    Article  PubMed  CAS  Google Scholar 

  42. Cardenas C, Quesada AR, Medina MA (2011) Anti-angiogenic and anti-inflammatory properties of kahweol, a coffee diterpene. PLoS One 6:e23407

    Article  PubMed  CAS  Google Scholar 

  43. Hai T, Hartman MG (2001) The molecular biology and nomenclature of the activating transcription factor/cAMP responsive element binding family of transcription factors: activating transcription factor proteins and homeostasis. Gene 273:1–11

    Article  PubMed  CAS  Google Scholar 

  44. Hai T, Wolfgang CD, Marsee DK, Allen AE, Sivaprasad U (1999) ATF3 and stress responses. Gene Expr 7:321–335

    PubMed  CAS  Google Scholar 

  45. Aung HH, Lame MW, Gohil K, He G, Denison MS, Rutledge JC et al (2011) Comparative gene responses to collected ambient particles in vitro: endothelial responses. Physiol Genomics 43:917–929

    Article  PubMed  CAS  Google Scholar 

  46. Vladykovskaya E, Sithu SD, Haberzettl P, Wickramasinghe NS, Merchant ML, Hill BG et al (2012) Lipid peroxidation product 4-hydroxy-trans-2-nonenal causes endothelial activation by inducing endoplasmic reticulum stress. J Biol Chem 287:11398–11409

    Article  PubMed  CAS  Google Scholar 

  47. Mungrue IN, Pagnon J, Kohannim O, Gargalovic PS, Lusis AJ (2009) CHAC1/MGC4504 is a novel proapoptotic component of the unfolded protein response, downstream of the ATF4-ATF3-CHOP cascade. J Immunol 182:466–476

    PubMed  CAS  Google Scholar 

  48. Turchi L, Aberdam E, Mazure N, Pouyssegur J, Deckert M, Kitajima S et al (2008) Hif-2alpha mediates UV-induced apoptosis through a novel ATF3-dependent death pathway. Cell Death Differ 15:1472–1480

    Article  PubMed  CAS  Google Scholar 

  49. Zhang C, Kawauchi J, Adachi MT, Hashimoto Y, Oshiro S, Aso T et al (2001) Activation of JNK and transcriptional repressor ATF3/LRF1 through the IRE1/TRAF2 pathway is implicated in human vascular endothelial cell death by homocysteine. Biochem Biophys Res Commun 289:718–724

    Article  PubMed  CAS  Google Scholar 

  50. Spinardi L, Einheber S, Cullen T, Milner TA, Giancotti FG (1995) A recombinant tail-less integrin beta 4 subunit disrupts hemidesmosomes, but does not suppress alpha 6 beta 4-mediated cell adhesion to laminins. J Cell Biol 129:473–487

    Article  PubMed  CAS  Google Scholar 

  51. Werner ME, Chen F, Moyano JV, Yehiely F, Jones JC, Cryns VL (2007) Caspase proteolysis of the integrin beta4 subunit disrupts hemidesmosome assembly, promotes apoptosis, and inhibits cell migration. J Biol Chem 282:5560–5569

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The work was supported in part by the National Natural Science Foundation of China (Nos. 90813022, 20972088, 31070735, 31000510 and 81021001), the National 973 Research Project (No. 2011CB503906), and the Natural Science Foundation of Shandong Province (ZR2010CZ004). We thank Dr. Laura Smales for the paper revising.

Author information

Authors and Affiliations

  1. Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, Jinan, 250100, China

    Di Ge, Xiangqian Kong, Jing Zhao, Le Su, Shangli Zhang & Junying Miao

  2. Institute of Organic Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan, 250100, China

    Weiyong Liu & Baoxiang Zhao

  3. The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Shandong University Qilu Hospital, Jinan, 250012, China

    Yun Zhang & Junying Miao

Authors
  1. Di Ge
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiangqian Kong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Weiyong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jing Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Le Su
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shangli Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yun Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Baoxiang Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Junying Miao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Baoxiang Zhao or Junying Miao.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 33 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ge, D., Kong, X., Liu, W. et al. Phosphorylation and nuclear translocation of integrin β4 induced by a chemical small molecule contribute to apoptosis in vascular endothelial cells. Apoptosis 18, 1120–1131 (2013). https://doi.org/10.1007/s10495-013-0860-4

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10495-013-0860-4

Keywords

Search

Navigation