, Volume 10, Issue 5, pp 963–971 | Cite as

Humanin delays apoptosis in K562 cells by downregulation of P38 MAP kinase

  • D. Wang
  • H. Li
  • H. Yuan
  • M. Zheng
  • C. Bai
  • L. Chen
  • X. Pei


Humanin (HN) is a newly identified neuroprotective peptide. In this study, we investigated its antiapoptotic effect and the potential mechanisms in K562 cells. Upon serum deprivation, expression of HN in K562 cells decreased and its intracellular distribution changed from cytoplasm to cell membrane. In HN stably transfected K562 cells, apoptosis was delayed compared with control vector transfected cells as measured by flow cytometry. Furthermore, analysis of different mitogen-activated protein (MAP) kinases activity revealed that extracellular signal-regulated kinase (ERK) pathway was inhibited while p38 signaling was activated following serum deprivation in K562 cells. And in HN transfected K562 cells, ERK downregulation was not affected, but p38 activation was suppressed, which may responsible for the delayed apoptosis in these cells. Activation of the ERK signaling pathway by phorbol myristate 13-acetate (PMA) and sorbitol protected K562 cells from serum deprivation induced apoptosis. Additionally, overexpression of HN reduced megakaryocytic differentiation of K562 cells. The present data outline the role of ERK and p38 MAP kinases in serum deprivation induced apoptosis in K562 cells and figure out p38 signaling pathway as molecular target for HN delaying apoptosis in K562 cells.


apoptosis differentiation humanin K562 cells MAPK 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Hashimoto Y, Niikura T, Tajima H, et al.. A rescue factor abolishing neuronal cell death by a wide spectrum of familial Alzheimer’s disease genes and AΒ. Proc Natl Acad Sci USA 2001; 98: 6336–6341.CrossRefPubMedGoogle Scholar
  2. 2.
    Hashimoto Y, Niikura T, Ito Y, et al.. Detailed characterization of neuroprotection by a rescue factor humanin against various Alzheimer’s disease-relevant insults. J Neurosci 2001; 21: 9235–9245.PubMedGoogle Scholar
  3. 3.
    Hashimoto Y, Ito Y, Niikura T, et al.. Mechanisms of neuroprotection by a novel rescue factor humanin from Swedish mutant amyloid precursor protein. Biochem Biophys Res Commun 2001; 283: 460–468.CrossRefPubMedGoogle Scholar
  4. 4.
    Kariya S, Takahashi N, Ooba N, Kawahara M, Nakayama H, Ueno S. Humanin inhibits cell death of serum-deprived PC12h cells. Neuroreport 2002; 13: 903–907.CrossRefPubMedGoogle Scholar
  5. 5.
    Mamiya T, Ukai M. [Gly(14)]-Humanin improved the learning and memory impairment induced by scopolamine in vivo. Br J Pharmacol 2001; 134: 1597–1599.CrossRefPubMedGoogle Scholar
  6. 6.
    Tajima H, Kawasumi M, Chiba T, et al.. A Humanin derivative, S14G-HN, prevents Amyloid-Β-induced memory impairment in mice. J Neurosci Res 2005; 79: 714–723.CrossRefPubMedGoogle Scholar
  7. 7.
    Jung SS, Van Nostrand WE. Humanin rescues human cerebrovascular smooth muscle cells from AΒ-induced toxicity. J Neurochem 2003; 84: 266–272.CrossRefPubMedGoogle Scholar
  8. 8.
    Ying G, Iribarren P, Zhou Y, et al.. Humanin, a newly identified neuroprotective factor, uses the G protein-coupled formylpeptide receptor-like-1 as a functional receptor. J Immunol 2004; 172: 7078–7085.PubMedGoogle Scholar
  9. 9.
    Hashimoto Y, Tsuji O, Niikura T, et al.. Involvement of c-Jun N-terminal kinase in amyloid precursor protein-mediated neuronal cell death. J Neurochem 2003; 84: 864–877.CrossRefPubMedGoogle Scholar
  10. 10.
    Guo B, Zhai DY, Gabezas E, et al.. Humanin peptide suppresses apoptosis by interfering with Bax activation. Nature 2003; 423: 456–461.CrossRefPubMedGoogle Scholar
  11. 11.
    Zhai D, Luciano F, Zhu X, Guo B, Satterthwait AC, Reed JC. Humanin binds and nullifies Bid activity by blocking its activation of Bax and Bak. J Biol Chem 2005; AOP.Google Scholar
  12. 12.
    Luciano F, Zhai D, Zhu X, et al.. Cytoprotective peptide Humanin binds and inhibits pro-apoptotic Bcl-2/Bax-family protein BimEL. J Biol Chem 2005; AOP.Google Scholar
  13. 13.
    Chang L, Karin M. Mammalian MAP kinase signaling cascades. Nature 2001; 410: 37–40.CrossRefPubMedGoogle Scholar
  14. 14.
    Witt O, Sand K, Pekrun A. Butyrate-induced erythroid differentiation of human K562 leukemia cells involves inhibition of ERK and activation of p38 MAP kinase pathways. Blood 2000; 95: 2391–2396.PubMedGoogle Scholar
  15. 15.
    Mayer IA, Verma A, Grumbach IM, et al..u The p38 MAPK pathway mediates the growth inhibitory effects of interferon-Ά in BCR-ABL-expressing cells. J Biol Chem 2001; 276: 28570–28577.CrossRefPubMedGoogle Scholar
  16. 16.
    Davis RJ. Signal transduction by the JNK group of MAP kinases. Cell 2000; 103: 239–252.CrossRefPubMedGoogle Scholar
  17. 17.
    McGee MM, Campiani G, Ramunno A, et al.. Activation of the c-Jun N-terminal kinase (JNK) signaling pathway is essential during PBOX-6-induced apoptosis in chronic myelogenous leukemia (CML) cells. J Biol Chem 2002; 277: 18383–18389.CrossRefPubMedGoogle Scholar
  18. 18.
    Zanjani ED, Almeida-Porada G, Livingston AG, Flake AW, Ogawa M. Human bone marrow CD34- cells engraft in vivo and undergo multilineage expression that includes giving rise to CD34+ cells. Exp Hematol 1998; 26: 353–360.PubMedGoogle Scholar
  19. 19.
    Bhatia M, Bonnet D, Murdoch B. A newly discovered class of human hematopoietic cells with SCID-repopulating activity. Nat Med 1998; 4: 1038–1045.CrossRefPubMedGoogle Scholar
  20. 20.
    Wang D, Yang L, Li L, et al.. Identification of differentially expressed genes in Lin-CD34- and Lin-CD34+ cells. Chin J Hematol 2003; 24: 423–425.Google Scholar
  21. 21.
    Kang CD, Yoo SD, Hwang BW, et al.. The inhibition of ERK/MAPK not the activation of JNK/SAPK is primarily required to induce apoptosis in chronic myelogenous leukemic K562 cells. Leukemia Res 2000; 24: 527–534.CrossRefGoogle Scholar
  22. 22.
    Kim DS, Hwang ES, Lee JE, Kim SY, Park KC. Sphingosine-1-phosphate promotes mouse melanocyte survival via ERK and Akt activation. Cell Signal 2003; 15: 919–926.CrossRefPubMedGoogle Scholar
  23. 23.
    Racke FK, Lewandowska K, Goueli S, Goldfarb AN. Sustained activation of the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway is required for megakaryocytic differentiation of K562 cells. J Biol Chem 1997; 272: 23366–23370.CrossRefPubMedGoogle Scholar
  24. 24.
    Goldfarb AN, Delehanty LL, Wang D, Racke FK, Hussaini IM. Stromal inhibition of megakaryocytic differentiation correlates with blockade of signaling by protein kinase C-ɛ and ERK/MAPK. J Biol Chem 2001; 276: 29526–29530.CrossRefPubMedGoogle Scholar
  25. 25.
    Sponne I, Fifre A, Koziel V, Kriem B, Oster T, Pillot T. Humanin rescues cortical neurons from prion-peptide-induced apoptosis. Mol Cell Neurosci 2004; 25: 95–102.PubMedGoogle Scholar
  26. 26.
    Brown JM, Attardi LD. The role of apoptosis in cancer development and treatment response. Nat Rev Cancer 2005; 5: 231–237.CrossRefPubMedGoogle Scholar
  27. 27.
    Miyoshi N, Uchida K, Osawa T, Nakamura Y. A link between benzyl isothiocyanate-induced cell cycle arrest and apoptosis: Involvement of mitogen-activated protein kinases in the Bcl-2 phosphorylation. Cancer Res 2004; 64: 2134–2142.PubMedGoogle Scholar
  28. 28.
    Lavelle D, DeSimone J, Hankewych M, Kousnetzova T, Chen YH. Decitabine induces cell cycle arrest at the G1 phase via p21 (WAF1) and the G2/M phase via the p38 MAP kinase pathway. Leuk Res 2003; 27: 999–1007.CrossRefPubMedGoogle Scholar
  29. 29.
    Desbarats J, Birge RB, Mimouni-Rongy M, Weinstein DE, Palerme JS, Newell MK. Fas engagement induces neurite growth through ERK activation and p35 upregulation. Nat Cell Biol 2003; 5: 118–125.CrossRefPubMedGoogle Scholar
  30. 30.
    Blalock WL, Navolanic PM, Steelman LS, et al.. Requirement for the PI3K/Akt pathway in MEK1-mediated growth and prevention of apoptosis: Identification of an Achilles heel in leukemia. Leukemia 2003; 17: 1058–1067.CrossRefPubMedGoogle Scholar
  31. 31.
    Aisa Y, Miyakawa Y, Nakazato T, et al.. Fucoidan induces apoptosis of human HS-sultan cells accompanied by activation of caspase-3 and downregulation of ERK pathways. Am J Hematol 2005; 78: 7–14.CrossRefPubMedGoogle Scholar
  32. 32.
    Lunghi P, Tabilio A, Dall’Aglio PP, et al.. Downmodulation of ERK activity inhibits the proliferation and induces the apoptosis of primary acute myelogenous leukemia blasts. Leukemia 2003; 17: 1783–1793.CrossRefPubMedGoogle Scholar
  33. 33.
    Planey SL, Abrams MT, Robertson NM, Litwack G. Role of apical caspases and glucocorticoid-regulated genes in glucocorticoid-induced apoptosis of pre-B leukemic cells. Cancer Res 2003; 63: 172–178.PubMedGoogle Scholar
  34. 34.
    Willaime-Morawek S, Brami-Cherrier K, Mariani J, Caboche J, Brugg B. C-Jun N-terminal kinases/c-Jun and p38 pathways cooperate in ceramide-induced neuronal apoptosis. Neuroscience 2003; 119: 387–397.CrossRefPubMedGoogle Scholar
  35. 35.
    Shelly C, Petruzzelli L, Herrera R. K562 cells resistant to phorbol 12-myristate 13-acetate-induced growth arrest: Dissociation of mitogen-activated protein kinase activation and Egr-1 expression from megakaryocyte differentiation. Cell Growth Differ 2000; 11: 501–506.PubMedGoogle Scholar
  36. 36.
    Sorrentino BP. Clinical strategies for expansion of haematopoietic stem cells. Nat Rev Immunol 2004; 4: 878–888.CrossRefPubMedGoogle Scholar
  37. 37.
    Domen J, Weissman IL. Hematopoietic stem cells need two signals to prevent apoptosis; Bcl-2 can provide one of these, Kitl/c-Kit signaling the other. J Exp Med 2000; 192: 1707–1718.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • D. Wang
    • 1
  • H. Li
    • 1
  • H. Yuan
    • 1
  • M. Zheng
    • 1
  • C. Bai
    • 1
  • L. Chen
    • 1
  • X. Pei
    • 1
  1. 1.Lab of Stem Cell BiologyBeijing Institute of Transfusion MedicineBeijingP. R. China

Personalised recommendations