Animal cells and systems

The Korean Society for Integrative Biology (한국통합생물학회)
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Caspase-11 Promoter-GFP Construct as a Dual Reporter of Cytotoxicity and Inflammation

  • Shin, Ki-Soon (Department of Biology, College of Sciences, Kyung Hee University) ;
  • Kang, Shin-Jung (Department of Molecular Biology, College of Natural Sciences, Sejong University)
  • Published : 2006.06.30
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Abstract

Caspase-11 has been known as a dual regulator of apoptosis and inflammatory response. An unusual feature of caspase-11 is that its expression is induced by apoptotic or proinflammatory stimuli. Utilizing these unusual features of caspase-11, we have developed a simple and sensitive assay method to screen pro- or anti-apoptotic/inflammatory molecules. To develop this assay method, we generated a reporter construct where GFP expression is regulated by caspase-11 promoter. When several types of cultured cells were transfected with this reporter construct and subsequently treated with various apoptotic or proinflammatory molecules, expression of GFP by the activation of caspase-11 promoter was easily detected by fluorescence microscopy or spectrofluorometry. In addition, a reduction of the GFP fluorescence was detected when an agent reported to suppress caspase-11 induction was applied. These results suggest that our reporter system can be used to screen pro- or anti-apoptotic/inflammatory molecules.

Keywords

References

  1. Ahn EY, Pan Q Vickers SM, and McDonald J. (2002) IFN-gamma upregulates apoptosis-related molecules and enhances Fas-mediated apoptosis in human cholangiocarcinoma. Int J Cancer 100: 445-451 https://doi.org/10.1002/ijc.10516
  2. Cryns V and Yuan J (1998) Proteases to die for. Genes Dev 12: 1551-1570 https://doi.org/10.1101/gad.12.11.1551
  3. Furuya T, Hayakawa H, Yamada M, Yoshimi K, Hisahara S, Miura M, Mizuno Y, and Mochizuki H (2004) Caspase-11 mediates inflammatory dopaminergic cell death in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson's disease. J Neuroci 24: 1865-1872
  4. Gao Z, Huang K, Yang X, and Xu H (1999) Free radical scavenging and antioxidant activities of flavonoids extracted from the radix of Scutellaria baicaensis. Georgi Biochirn Biophys Acta 1472: 643-650 https://doi.org/10.1016/S0304-4165(99)00152-X
  5. Hisahara S, Yuan J, Momoi T, Okano H, and Miura M (2001) Caspase-11 mediates oligodendrocyte cell death and pathogenesis of autoimmune-mediated demyelination. J Exp Med 193: 111-122 https://doi.org/10.1084/jem.193.1.111
  6. Hu P, Han Z, Couvillon AD, and Exton JH (2004) Critical role of endogenous AKT/IAPs and MEK1/ERK pathways in counteracting endoplasmic reticulum stress-induced cell death. J Bioi Chem 279: 49420-49429 https://doi.org/10.1074/jbc.M407700200
  7. Jaeschke H (2002) Xanthine oxidase-induced oxidant stress during hepatic ischemia-reperfusion: are we coming full circle after 20 years? Hepatology 36: 761-763 https://doi.org/10.1053/jhep.2002.36038
  8. Kang K, Oh YK, Choue R, and Kang SJ (2005) Scutellariae radix extracts suppress ethanol-induced caspase-11 expression and cell death in N2a cells. Brain Res Mol Brain Res 142: 139-145 https://doi.org/10.1016/j.molbrainres.2005.09.006
  9. Kang SJ, Wang S, Hara H, Peterson EP, Namura S, Amin-Hanjani S, Huang Z, Srinivasan A, Tomaselli KJ, Thornberry NA, Moskowitz MA, and Yuan J (2000) Dual role of caspase-11 in mediating activation of caspase-1 and caspase- 3 under pathological conditions. J Cell Biol 149: 613-622 https://doi.org/10.1083/jcb.149.3.613
  10. Kang SJ, Wang S, Kuida K, and Yuan J (2002) Distinct downstream pathways of caspase-11 in regulating apoptosis and cytokine maturation during septic shock response. Cell Death Differ 9: 1115-1125 https://doi.org/10.1038/sj.cdd.4401087
  11. Lin XY, Choi MS, and Poter AG (2000) Expression analysis of the human caspase-1 subfamily reveals specific regulation of the CASP5 gene by lipopolysaccharide and interferon-gamma. J Bioi Chem 275: 39920-39926 https://doi.org/10.1074/jbc.M007255200
  12. Martinon F, Bums K, and Tschopp J (2002) The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proIL-beta. Mol Cell 10: 417-426 https://doi.org/10.1016/S1097-2765(02)00599-3
  13. McCarthy KD, and de Vellies J (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85: 890-902 https://doi.org/10.1083/jcb.85.3.890
  14. Olney JW, Womiak DF, Farber NB, Jevtovic-Todorovic V, Bittigau P, and Ikonomidou C (2002) The enigma of fetal alcohol neurotoxicity. Ann Med 34: 109-119 https://doi.org/10.1080/07853890252953509
  15. Phillips A, Bullock T, and Plant N (2003) Sodium valproate induces apoptosis in the rat hepatoma cell line, FaO. Toxicology 192: 219-227 https://doi.org/10.1016/S0300-483X(03)00331-7
  16. Schauvliege R, Vanrobaeys J, Schotte P, and Beyaert R (2002) Caspase-11 gene expression in response to lipopolysaccharide and interferon-gamma requires nuclear factor-kappa B and signal transducer and activator of transcription (STAT) 1. J Biol Chem 277: 41624-41630 https://doi.org/10.1074/jbc.M207852200
  17. ShiehDE, Liu LT, and Lin CC (2000) Antioxidant and free radical scavenging effects of baicalein, baicalin and wogonin. Anticancer Res 20: 2861-2865
  18. Suk K, Lee H, Kang SS, Cho GJ, and Choi WS (2003) Flavonoid baicalein attenuates activation-induced cell death of brain microglia. J Pharmacol Exp Ther 305: 638-645 https://doi.org/10.1124/jpet.102.047373
  19. Wang M, Tsai BM, Kher A, Baker LB, Wairiuko GM, and Meldrum DR (2005) Role of endogenous testosterone in myocardial proinflammatory and proapoptotic signaling after acute ischemia-reperfusion. Am J Physiol Heart Circ Physiol 288: H221-226 https://doi.org/10.1152/ajpheart.00784.2004
  20. Wang S, Miura M, Jung Y, Zhu H, Gagliardini V, Shi L, Greenberg AH, and Yuan J (1996) Identification and characterization of Ich-3, a member of the interleukin-lbeta converting enzyme (lCE)/Ced-3 family and an upstream regulator of ICE. J Biol Chem 271: 20580-20587 https://doi.org/10.1074/jbc.271.34.20580
  21. Wang S, Miura M, Jung YK, Zhu H, Li E, and Yuan J (1998) Murine caspase-11, an ICE-interacting protease, is essential for the activation of ICE. Cell 92: 501-509 https://doi.org/10.1016/S0092-8674(00)80943-5
  22. Yoon BI, Li GX, Kitada K, Kawasaki Y, Igarashi K, Kodama Y, Inoue T, Kobayashi K, Kanno J, Kim DY, Inoue T, and Hirabayashi Y (2003) Mechanisms of benzene-induced hematotoxicity and leukemogenicity: cDNA microarray analyses using mouse bone marrow tissue. Environ Health Perspect 111: 1411-1420 https://doi.org/10.1289/ehp.6164
  23. Kobori M, Yang Z, Gong D, Heissmeyer V, Zhu H, Jung YK, Gakidis MA, Rao A, Sekine T, Ikegami F, Yuan C, and Yuan J (2004) Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK complex. Cell Death Differ 11: 123-30 https://doi.org/10.1038/sj.cdd.4401325