Cytotoxicological Effect of Tebufenozide, an Insect Growth Regulator (IGR): Stimulation of Filamentous Actin Reorganization and Enhancement of Hsp27 Expression in Drosophila Kc Cells

  • Hwang, Jee-Na (Department of Pharmacology, Medical Research Center (MRC), Kyung Hee University School of Medicine) ;
  • Jung, Hwa-Jin (Department of Pharmacology, Medical Research Center (MRC), Kyung Hee University School of Medicine) ;
  • Seo, Young-Rok (Department of Pharmacology, Medical Research Center (MRC), Kyung Hee University School of Medicine)
  • Published : 2004.12.21

Abstract

The cytotoxicological responses to insect growth regulator (IGR), using tebufenozide as ecdysteroid mimic, were investigated in Drosophila Kc cells. Treatment of Kc cells with tebufenozide showed significant growth inhibition and striking morphological changes including aggregation and elongation of the cells. In order to understand the cellular mechanism underlying the response of Drosophila cells to tebufenozide, immunofluorescence microscopy was performed. We found that treatment of Kc cells with tebufenozide enhanced the reorganization of f-actin and stimulated the expression of hsp27. These data suggest a possible association of filamentous actin (f-actin) and hsp27 in the cytotoxicological mechanisms of growth regulators in Drosophila cells.

Keywords

References

  1. Anne JM, Martin SJ, Bissonnette RP, Mahboubi A, Shi Y, Mogil RJ, Nishioka WK, Green DR. Cell Death. In: Schwartz L, Osborne B ed, Methods in Cell Biology. Academic Press, San Diego, CA, p 153-185, 1995
  2. Best-Belpomme M, Courgeon AM, Rambach A. Galactosidase is induced by hormone in Drosophila melanogaster cell cultures. Proc Natl Acad Sci USA 75: 6102-6106, 1978 https://doi.org/10.1073/pnas.75.12.6102
  3. Chen L, Reece C, O'Keefe SL, Hawryluk GW, Engstrom MM, Hodgetts RB. Induction of the early-late Ddc gene during Drosophila metamorphosis by the ecdysone receptor. Mech Dev 114: 95-107, 2002 https://doi.org/10.1016/S0925-4773(02)00064-3
  4. Chen GC, Gajowniczek P, Settleman J. Rho-LIM kinase signaling regulates ecdysone-induced gene expression and morphogenesis during Drosophila metamorphosis. Curr Biol 14: 309-313, 2004 https://doi.org/10.1016/j.cub.2004.01.056
  5. Cherbas L, Bene H, Bourouis M, Burtis K, Chao A, Cherbas P, Crosby M, Garfinkel M, Guild G, Hogness D, Jami J, Jones CW, Koehler M, Lepesant JA, Martin C, Maschat F, Mathers P, Meyerowitz E, Moss R, Pictet R, Rebers J, Richards G, Roux J, Schulz R, Segraves W, Thummel C, Vijayraghavan K. Structural and functional analysis of some moulting hormone-responsive genes from Drosophila. Insect Biochem 16: 241-248, 1986 https://doi.org/10.1016/0020-1790(86)90102-2
  6. Cherbas L, Fristrom JW and O'connor JD. Biosynthesis, Metabolism and Mode of Action of Invertebrate Hormones. In: Hoffmann J, Porchet M ed, Proceedings in Life Sciences. Springer- Verlag, Berlin, p 305-322, 1984
  7. Cherbas P, Cherbas L, Williams CM. Induction of acetylcholinesterase activity by -ecdysone in a Drosophila cell line. Science 197: 275-277, 1977 https://doi.org/10.1126/science.877552
  8. Courgeon AM. Action of insect hormones at the cellular level: Morphological changes of a diploid cell line of Drosophila melanogaster treated with ecdysone and several analogs in vitro. Exp Cell Res 74: 327-336, 1972 https://doi.org/10.1016/0014-4827(72)90384-9
  9. de Jong WW, Leunissen JA, Voorter CE. Evolution of the alphacrystallin/ small heat-shock protein family. Mol Biol Evol 10: 103 -126, 1993
  10. Echalier G, Ohanessian A. Isolation in tissue culture, of Drosophila melangaster cell lines. C R Acad Sci Hebd Seances Acad Sci D. 268: 1771-1773, 1969
  11. Geneviĕve M, Yutaka I, Kanefusa K, Robert MT. The small heat shock protein Hsp22 of Drosophila melanogaster is a mitochondrial protein displaying oligomeric organization. J Biol Chem 275: 31204-31210, 2000 https://doi.org/10.1074/jbc.M002960200
  12. Hock T, Cottrill T, Keegan J, Garza D. The E23 early gene of Drosophila encodes an ecdysone-inducible APT-binding cassette transporter capable of repressing ecdysone-mediated gene activation. PNAS 97: 9519-9524, 2000 https://doi.org/10.1073/pnas.160271797
  13. Li GC, Li L, Liu YK, Mak JY, Lee C. Thermal response of rat fibroblasts stably transfected with the human 70-kDa heat shock protein-encoding genes. Proc Natl Acad Sci USA 88: 1681-1685, 1991 https://doi.org/10.1073/pnas.88.5.1681
  14. Marks EP. Insect tissue culture: An overview. Annu Rev Entomol 73: 1971-1978, 1980
  15. Mehlen P, Kretz-Remy C, Preville X, Arrigo AP. Human hsp27, Drosophila hsp27 and human alphaB-crystallin expressionmediated increase in glutathione is essential for the protective activity of these proteins against TNF alpha-induced cell death. EMBO J 15: 2695-2706, 1996
  16. Nakagawa Y, Minakuchi C, Takahashi K, Ueno T. Inhibition of $[^3H]$ponasterone A binding by ecdysone agonists in the intact Kc cell line. Insect Biochem and Mol Biol 32: 175-180, 2002 https://doi.org/10.1016/S0965-1748(01)00105-9
  17. Riddiford LM. Atlas of Drospheila Development: Drosophila Third Instar Eye Disk Chart. In: Bate M, Arias AM ed, The development of Drosophila melanogaster. Cold Spring Horbor Lab Press, Plainview, NY, p 899-939, 1993
  18. Ritossa F. A New puffing pattern induced by temperature shock and DNP in Drosophila. Experientia 18: 571-573, 1962 https://doi.org/10.1007/BF02172188
  19. Ropp M, Courgeon AM, Calvayrac R, Best-Belpomme M. The possible role of the superoxide ion in the induction of heat-shock and specific proteins in aerobic Drosophila cells during return to normoxia after a period of anaerobiosis. Can J Biochem Cell Biol 61: 456-461, 1983 https://doi.org/10.1139/o83-061
  20. Wang HD, Kazemi-Esfarjani P, Benzer S. Multiple-stress analysis for isolation of Drosophila longevity genes. PNAS 101: 12610- 12615, 2004 https://doi.org/10.1073/pnas.0404648101