Regulation of Endothelin-Converting Enzyme–1 Expression in Human Neuroblastoma Cells

Abstract

In recent years endothelin-converting enzyme (ECE-1) has been suggested to play an important role in amyloid-β peptide metabolism as one of the amyloid-degrading enzymes. In this connection, the analysis of the levels of expression and distribution of ECE-1 in the brain under normal and pathologic conditions could be important in neurodegeneration and pathogenesis of Alzheimer disease. In our previous studies, we have demonstrated that expression of ECE-1 was significantly reduced in the cortex of adult rats after 15 mins of global ischemia. It was also significantly reduced in the striatum of rats subjected to prenatal hypoxia. In the present study, we analyzed effects of hypoxia and oxidative stress on ECE-1 in human neuroblastoma NB7 cells and effects of the cholinergic agonist carbachol and the phorbol ester, phorbol 12-myristate 13-acetate (PMA). We have found that chronic (24 hrs) hypoxia and oxidative stress resulted in 30% and 20% decrease in expression of ECE-1 at the protein level, respectively, although at the level of ECE-1 mRNA there were no statistically significant changes. Serum withdrawal from the incubation medium as well as addition of carbachol or PMA for 24 hrs also led to a significant reduction of the levels of ECE-1 protein in NB7 cells. Further study of the downstream signaling cascades involved in downregulation of ECE expression in NB7 cells and primary neuronal cells might provide us with new insights into possible therapeutic strategies for prevention or treatment of Alzheimer disease in elderly patients and those who suffer from stroke or cerebrovascular disorders.

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References

Xu D, Emoto N, Giaid A, Slaughter C, Kaw S, deWit D, Yanagisawa M. ECE-1: a membrane-bound metalloprotease that catalyzes the proteolytic activation of big endothelin-1. Cell 78:473–485, 1994.

Matsumura Y, Ikegawa R, Takaoka M, Morimoto S. Conversion of porcine big endothelin to endothelin by an extract from the porcine aortic endothelial cells. Biochem Biophys Res Commun 167:203–210, 1990.

Takahashi M, Matsushita Y, Iijima Y, Tanzawa K. Purification and characterization of endothelin converting enzyme from rat lung. J Biol Chem 268:21394–21398, 1993.

Takahashi M, Fukuda K, Shimada K, Barnes K, Turner AJ, Ikeda M, Koike M, Yamamoto Y, Tanzawa K. Localization of rat endothelin converting enzyme to vascular endothelial cells and some secretory cells. Biochem J 311:657–665, 1995.

Barnes K, Turner AJ. Endothelin converting enzyme is located on β-actin filaments in smooth muscle cells. Cardiovasc Res 42:814–822, 1999.

Barnes K, Walkden BJ, Wilkinson TC, Turner AJ. Expression of endothelin-converting enzyme in both neuroblastoma and glial cell lines and its localization in rat hippocampus. J Neurochem 68:570–577, 1997.

Schweizer A, Valdenaire O, Nelböck P, Deuschle U, Dumas M, Edwards JB, Stumpf JG, Löffler BM. Human endothelin converting enzyme (ECE-1): three isoforms with distinct subcellular localizations. Biochem J 328:871–877, 1997.

Valdenaire O, Lepailleur-Enouf D, Edidy G, Thouard A, Barrer A, Vranckx R, Tougard C, Michel JB. A fourth isoform of endothelin-converting enzyme (ECE-1) is generated from an additional promoter. Eur J Biochem 264:341–349, 1999.

Turner AJ, Barnes K, Schweizer A, Valdenaire O. Isoforms of endothelin converting enzyme: why and where? Trends Pharmacol Sci 19:483–486, 1998.

10.

Hoang MV, Turner AJ. Novel activity for endothelin-converting enzyme: hydrolysis of bradykinin. Biochem J 327:23–26, 1997.

11.

Johnson GD, Stevenson T, Ahn K. Hydrolysis of peptide hormones by endothelin-converting enzyme-1. A comparison with neprilysin. J Biol Chem 274:4053–4058, 1999.

12.

Eckman EA, Reed DK, Eckman CB. Degradation of the Alzheimer’s amyloid β peptide by endothelin-converting enzyme. J Biol Chem 276:24540–24548, 2001.

13.

Hofman A, Ott A, Breteler MM, Bots ML, Slooter AJ, Van Harskamp F, van Duijn CN, Van Broeckhoven C, Grobbee DE. Atherosclerosis, apolipoprotein E, and prevalence of dementia and Alzheimer’s disease in the Rotterdam Study. Lancet 349:151–154, 1997.

14.

Nalivaevaa NN, Fisk L, Kochkina EG, Plesneva SA, Zhuravin IA, Babusikova E, Dobrota D, Turner AJ. Effect of hypoxia/ischemia and hypoxic preconditioning/reperfusion on expression of some amyloid-degrading enzymes. Ann N Y Acad Sci 1035:21–33, 2004.

15.

Nalivaeva NN, Fisk L, Canet Aviles RM, Plesneva SA, Zhuravin IA, Turner AJ. Effects of prenatal hypoxia on expression of amyloid precursor protein and metallopeptidases in the rat brain. Lett Peptide Sci 10:455–462, 2003.

16.

Canet-Aviles RM, Anderton M, Hooper NM, Turner AJ, Vaughan PF. Muscarine enhances soluble amyloid precursor protein secretion in human neuroblastoma SH-SY5Y by a pathway dependent on protein kinase C(α), src-tyrosine kinase and extracellular signal-regulated kinase but not phospholipase C. Brain Res Mol Brain Res 102:62–72, 2002.

17.

Zhu G, Wang D, Lin YH, McMahon T, Koo EH, Messing RO. Protein kinase Cε suppresses Aβ production and promotes activation of α-secretase. Biochem Biophys Res Commun 285:997–1006, 2001.

18.

Brown CD, Barnes K, Turner AJ. Anti-peptide antibodies specific to rat endothelin-converting enzyme-1 isoforms reveal isoform localisation and expression. FEBS Lett 424:183–187, 1998.

19.

Zhang JZ, Behrooz A, Ismail-Beigi F. Regulation of glucose transport by hypoxia. Am J Kidney Dis 34:189–202, 1999.

20.

Evangelopoulos ME, Weis J, Kruttgen A. Signalling pathways leading to neuroblastoma differentiation after serum withdrawal: HDL blocks neuroblastoma differentiation by inhibition of EGFR. Oncogene 24:3309–3318, 2005.

21.

Nalivaeva NN, Turner AJ. Does acetylcholinesterase secretion involve an ADAMS-like metallosecretase? Lett Peptide Sci 6:343–348, 1999.

22.

Orzechowski HD, Gunther A, Menzel S, Zimmermann A, Funke-Kaiser H, Real R, Subkowski T, Zollmann FS, Paul M. Transcriptional mechanism of protein kinase C-induced isoform-specific expression of the gene for endothelin-converting enzyme-1 in human endothelial cells. Mol Pharmacol 60:1332–1342, 2001.

23.

Oh-hashi K, Nagai T, Tanaka T, Yu H, Hirata Y, Kiuchi K. Determination of hypoxic effect on neprilysin activity in human neuroblastoma SH-SY5Y cells using a novel HPLC method. Biochem Biophys Res Commun 334:380–385, 2005.

24.

Berger Y, Dehmlow H, Blum-Kaelin D, Kitas EA, Loffler BM, Aebi JD, Juillerat-Jeanneret L. Endothelin-converting enzyme-1 inhibition and growth of human glioblastoma cells. J Med Chem 48:483–498, 2005.

25.

Jolly-Tornetta C, Wolf BA. Regulation of amyloid precursor protein (APP) secretion by protein kinase Cα in human ntera 2 neurons (NT2N). Biochemistry 39:7428–7435, 2000.

26.

Geiger M, Wrulich OA, Jenny M, Schwaiger W, Grunicke HH, Uberall F. Defining the human targets of phorbol ester and diacylglycerol. Curr Opin Mol Ther 6:631–641, 2003.