Apoptosis-Induced Gene Profiles of a Myeloma Cell P3-X63-Ag8.653

  • Bahng, Hye-Seung (Department of Microbiology, Hanyang University College of Medicine) ;
  • Chung, Yong-Hoon (Department of Microbiology, College of Medicine, Hanyang University)
  • Published : 2006.09.30

Abstract

Background: Apoptosis is a physiologic phenomenon involved in development, elimination of damaged cells, and maintenance of cell homeostasis. Deregulation of apoptosis may cause diseases, such as cancers, immune diseases, and neurodegenerative disorders. The mouse myeloma cell P3-X63-Ag8.653 (v653) is an HGPRT deficient $(HGPRT^-)$ mutant strain. High dependency on de novo transcription and translation of aminopterin induced apoptosis of this cell seems to be an ideal experimental system for searching apoptosis-induced genes. Methods & Results: For searching apoptosis-related genes we carried out GE-array (dot blot), Affymetrix GeneChip analysis, Northern analysis and differential display-PCR techniques. The chip data were analyzed with three different programs. 66 genes were selected through Affymetrix GeneChip analyses. All genes selected were classified into 8 groups according to their known functions. They were Genes of 1) Cell growth/maintenance/death/enzyme, 2) Cell cycle, 3) Chaperone, 4) Cancer/disease-related genes, 5) Mitochondria, 6) Membrane protein/signal transduction, 7) Nuclear protein/nucleic acid binding/transcription binding and 8) Translation factor. Among these groups number of genes were the largest in the genes of cell growth/maintenance/death/enzyme. Expression signals of most of all groups were peaked at 3 hour of apoptosis except genes of Nuclear protein/nucleic acid binding/transcription factor which showed maximum signal at 1 hour. Conclusion: This study showed induction of wide range of proapoptotic factors which accelerate cell death at various stage of cell death. In addition apoptosis studied in this research can be classified as a type 2 which involves cytochrome c and caspase 9 especially in early stages of death. But It also has progressed to type 1 in late stage of the death process.

Keywords

References

  1. Kerr JF, Wyllie AH, Currie AR: Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26;239-257, 1972 https://doi.org/10.1038/bjc.1972.33
  2. Kearney JF, Radbruch A, Liesegang B, Rajesky K: A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol 123;1548-1550, 1979
  3. Abbas A, Lightman AH, Pober J. In: WTonsiewicz MJ ed.: Cellular and Molecular Immunology Oncology, p37-68, Saunders, London, 1990
  4. Zacharchuk CM, Mercep M, Chakraborti PK, Simons SS, Ashwell JD: Programmed T lymphocyte death. Cell activation-and steroid-induced pathways are mutually antagonistic. J. Immunol 145;4037-4045, 1990
  5. Chung Y, Youn J, Choi Y, Paik D, Cho Y: Requirement of de novo protein synthesis for aminopterin-induced apoptosis in a mouse myeloma cell line. Immunol Letters 77;127-131, 2001 https://doi.org/10.1016/S0165-2478(01)00213-9
  6. Sakamuro D, Eviner V, Elliott KJ, Showe L, White E, Prendergast GC: c-myc induces apoptosis in epithelial cells by both p53-dependent and p53-independent mechanisms. Oncogene 11;2411-2418, 1995
  7. Muga SJ, Grider A: Partial characterization of a human zincdeficiency syndrome by differential display. Biol Trace Elem Res 68;1-12, 1999 https://doi.org/10.1007/BF02784392
  8. Stegmaier K, Ross KN, Colavito SA, O'Malley S, Stockwell BR, Golub TR: Gene expression-based high-throughput screening (GE-HTS) and application to leukemia differentiation. Nat Genet 36;257-263, 2004 https://doi.org/10.1038/ng1305
  9. Wilson JW, Bean P, Robins T, Graziano F, Persing DH: Comparative evaluation of three human immunodeficiency virus genotyping systems: the HIV-GenotypR method, the HIV PRT GeneChip assay and the HIV-1 RT line probe assay. J Clin Microbiol 38;3022-3028, 2000
  10. Huang GS, Hong MY, Liu YC: Incorporation of DNA chip technology to the simulation and validation of flux analysis in yeast diauxic growth. Life Sci 72;2525-2531, 2003 https://doi.org/10.1016/S0024-3205(03)00138-3
  11. Dahlquist KD, Salomonis N, Vranizan K, Lawlor SC, Conklin BR: GenNIAPP, a new tool for viewing and analyzing micro array data on biological pathways. Nat Genet 31 ;19-20, 2002 https://doi.org/10.1038/ng0502-19
  12. Lisi S, Sisto M, Acquafredda A, Spinelli R, Schiavone M, lVIitolo V, Brandonisio O, Panaro M: Infection with Leishmania infantum inhibits actinomycin D-induced apoptosis of human monocytic cell line U-937. J Eukaryot Microbiol 52;211-217, 2005 https://doi.org/10.1111/j.1550-7408.2005.00026.x
  13. Fraschini A, Bottone MG, Scovassi AI, Denegri M, Risueno MC, Testillano PS, Martin TE, Biggiogera M, Pellicciari C: Changes in extranucleolar transcription during actinomycin D-induced apoptosis. Histol Histopathol 20;107-117, 2005
  14. Kadenbach B, Arnold S, Lee I, Huttemann M: The possible role of cytochrome c oxidase in stress-induced apoptosis and degenerative diseases. Biochim Biophys Acta 1655;400-408, 2004 https://doi.org/10.1016/j.bbabio.2003.06.005
  15. Burkitt M, Jones C, Lawrence A, Wardman P: Activation of cytochrome c to a peroxidase compound I-type intermediate by $H_{2}O_{2}$: relevance to redox signalling in apoptosis. Biochem Soc Symp 71 ;97 -106, 2004
  16. Jiang X, Wang X: Cytochrome C-mediated apoptosis. Annu Rev Biochem 73;87-106, 2004 https://doi.org/10.1146/annurev.biochem.73.011303.073706
  17. Kirkland RA, Franklin JL: Bax, reactive oxygen, and cytochrome c release in neuronal apoptosis. Antioxid Redox Signal 5;589-596, 2003 https://doi.org/10.1089/152308603770310257
  18. Ramadani M, Gansauge F, Schlosser S, Yang Y, Beger HG, Gansauge S: Overexpression of caspase-l in pancreatic disorders: implications for a function besides apoptosis. J Gastrointest Surg 5;352-358, 2001 https://doi.org/10.1016/S1091-255X(01)80061-5
  19. Castedo M, Perfettini JL, Roumier T, Krocmer G: Cyclindependent kinase-1: linking apoptosis to cell cycle and mitotic catastrophe. Cell Death Differ 9;1287-1293, 2002 https://doi.org/10.1038/sj.cdd.4401130
  20. Baar RA, Dingfelder CS, Smith LA, Bernlohr DA, Wu C, Lange AJ, Parks EJ: Investigation of in vivo fatty acid metabolism in AF ABP / aP2( - / -) mice. Am J Physiol Endocrinol Metab 288;187-193, 2005 https://doi.org/10.1152/ajpendo.00256.2004
  21. Boord JB, Maeda K, Makowski L, Babaev VR, Fazio S, Linton MF, Hotamisligil GS: Adipocyte fatty acid-binding protein, aP2, alters late atherosclerotic lesion formation in severe hypercholesterolemia. Arterioscler Thromb Vase Biol 22; 1686-1691, 2002 https://doi.org/10.1161/01.ATV.0000033090.81345.E6
  22. Maeda T, Hanna AN, Sim AB, Chua PP, Chong MT, Tron VA: GADD45 regulates G2/M arrest, DNA repair, and cell death in keratinocytes following ultraviolet exposure. Invest Dermatol 119;22-26, 2002 https://doi.org/10.1046/j.1523-1747.2002.01781.x
  23. Salvador JM, Hollander MC, Nguyen AT, Kopp JB, Barisoni L, Moore JK, Ashwell JD, Fornace AJ Jr: Mice lacking the p53-effector gene Gadd45a develop a lupus-like syndrome. Immunity 6;499-508, 2002