Advertisement

International Journal of Hematology

, Volume 79, Issue 3, pp 276–282 | Cite as

Monitoring the expression profiles of doxorubicin-resistant K562 human leukemia cells by serial analysis of gene expression

  • Yoshikazu Ichikawa
  • Makoto Hirokawa
  • Namiko Aiba
  • Naohito Fujishima
  • Atsushi omatsuda
  • Hirobumi Saitoh
  • Masaaki Kume
  • Ikuo Miura
  • Ken-ichi Sawada
Case Report

Abstract

We examined the expression profiles of doxorubicin-resistant K562 cells by serial analysis of gene expression (SAGE) to identify novel and/or partially characterized genes that might be related to drug resistance in human leukemia. SAGE complementary DNA (cDNA) libraries were constructed from K562 and doxorubicin-resistant K562 (K562/ADM) cells, and concatamer sequences were analyzed with SAGE 2000 software.We used 9792 tags in the identification of 1076 different transcripts, 296 of which were similarly expressed in K562 and K562/ADM cells.There were 343 genes more actively expressed in K562/ADM than in parental K562 cells and 437 genes expressed less often in K562/ADM cells. K562/ADM cells showed increased expression of well-known genes, including the genes for spectrin β, eukaryotic translation initiation factor 1A (EIF1A), RAD23 homolog B, laminin receptor 1, and polyA-, RAN-, and PAI-1 messenger RNA-binding proteins. K562/ ADM cells showed decreased expression of the genes for fatty acid desaturase 1 (FADS1), hemoglobin ε 1, N-myristoyltransferase 1, hemoglobin α 2, NADH dehydrogenase Fe-S protein 6, heat shock 90-kDa protein, and karyopherin β1. Quantitative reverse transcription-polymerase chain reaction analysis confirmed the increased expression of EIF1A and the decreased expression of FADS1 in K562/ADM cells. Prior to this investigation, such differences in the expression of these genes in doxorubicinresistant leukemia cells were unknown. Although we do not provide any evidence in the present report for the potential roles of these genes in drug resistance, SAGE may provide a perspective into our understanding of drug resistance in human leukemia that is different from that provided by cDNA microarray analysis.

Key words

SAGE Expression profile K562 Doxorubicin Resistance 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Fisher DE. Apoptosis in cancer therapy: crossing the threshold. Cell. 1994;78:539–542.CrossRefGoogle Scholar
  2. 2.
    Kawabata Y, Hirokawa M, Kitabayashi A, Horiuchi T, Kuroki J, Miura AB. Defective apoptotic signal transduction pathway downstream of caspase-3 in human B lymphoma cells: a novel mechanism of nuclear apoptosis resistance. Blood. 1999;94:3523–3530.Google Scholar
  3. 3.
    Dalton WS. Mechanisms of drug resistance in hematologic malignancies. Semin Hematol. 1997;34(suppl 5):3–8.Google Scholar
  4. 4.
    Hirokawa M, Kawabata Y, Miura AB. Dysregulation of apoptosis and a novel mechanism of defective apoptotic signal transduction in human B-cell neoplasms. Leuk Lymphoma. 2002;43:243–249.CrossRefGoogle Scholar
  5. 5.
    Kudoh K, Ramanna M, Ravatn R, et al. Monitoring the expression profiles of doxorubicin-induced and doxorubicin-resistant cancer cells by cDNA microarray. Cancer Res. 2000;60:4161–4166.Google Scholar
  6. 6.
    Turton NJ, Judah DJ, Riley J, et al. Gene expression and amplification in breast carcinoma cells with intrinsic and acquired doxorubicin resistance. Oncogene. 2001;20:1300–1306.CrossRefGoogle Scholar
  7. 7.
    Sakamoto M, Kondo A, Kawasaki K, et al. Analysis of gene expression profiles associated with cisplatin resistance in human ovarian cancer cell lines and tissues using cDNA microarray. Hum Cell. 2001;14:305–315.Google Scholar
  8. 8.
    Zembutsu H, Ohnishi Y, Tsunoda T, et al. Genome-wide cDNA microarray screening to correlate gene expression profiles with sensitivity of 85 human cancer xenografts to anticancer drugs. Cancer Res. 2002;62:518–527.Google Scholar
  9. 9.
    Kihara C, Tsunoda T, Tanaka T, et al. Prediction of sensitivity of esophageal tumors to adjuvant chemotherapy by cDNA microarray analysis of gene-expression profiles. Cancer Res. 2001;61: 6474–6479.Google Scholar
  10. 10.
    Velculescu VE, Zhang L, Vogelstein B, Kinzler KW. Serial analysis of gene expression. Science. 1995;270:484–487.CrossRefGoogle Scholar
  11. 11.
    Tsuruo T, Saito HI, Kawabata H, Oh-hara T, Hamada H, Utakoji T. Characteristics of resistance to Adriamycin in human myelogenous leukemia K562 resistant to Adriamycin and in isolated clones. Jpn J Cancer Res. 1986;77:682–692.Google Scholar
  12. 12.
    Sugimoto Y, Tsuruo T. DNA-mediated transfer and cloning of a human multidrug-resistant gene of Adriamycin-resistant myelogenous leukemia K562. Cancer Res. 1987;47:2620–2625.Google Scholar
  13. 13.
    Naito M, Tsuruo T. Competitive inhibition by verapamil of ATPdependent high affinity vincristine binding to the plasma membrane of multidrug-resistant K562 cells without calcium ion involvement. Cancer Res. 1989;49:1452–1455.Google Scholar
  14. 14.
    Sugawara I, Iwahashi T, Okamoto K, et al. Characterization of an etoposide-resistant human K562 cell line, K/eto. Jpn J Cancer Res. 1991;82:1035–1043.CrossRefPubMedGoogle Scholar
  15. 15.
    Batty D, Rapic’-Otrin V, Levine AS, Wood RD. Stable binding of human XPC complex to irradiated DNA confers strong discrimination for damaged sites. J Mol Biol. 2000;300:275–290.CrossRefGoogle Scholar
  16. 16.
    Sugasawa K, Ng JM, Masutani C, et al. Xeroderma pigmentosum group C protein complex is the initiator of global genome nucleotide excision repair. Mol Cell. 1998;2:223–232.CrossRefGoogle Scholar
  17. 17.
    Volker M, Mone MJ, Karmakar P, et al. Sequential assembly of the nucleotide excision repair factors in vivo. Mol Cell. 2001;8: 213–224.CrossRefGoogle Scholar
  18. 18.
    Tewely KM, Rowe TC, Yang L, Halligan BD, Liu LF. Adriamycininduced DNA damage mediated by mammalian DNA topoisomerase II. Science. 1984;226:466–468.CrossRefGoogle Scholar
  19. 19.
    Yow HK, Wong JM, Chen HS, et al. Increased mRNA expression of a laminin-binding protein in human colon carcinoma: complete sequence of a full-length cDNA encoding the protein. Proc Natl Acad Sci U S A. 1988;85:6394–6398.CrossRefPubMedGoogle Scholar
  20. 20.
    Damiano JS, Hazlehurst LA, Dalton WS. Cell adhesion-mediated drug resistance (CAM-DR) protects the K562 chronic myelogenous leukemia cell line from apoptosis induced by BCR/ABL inhibition, cytotoxic drugs, and gamma-irradiation. Leukemia. 2001;15: 1232–1239.CrossRefGoogle Scholar
  21. 21.
    Shi Y, Zhai H, Wang X, et al. Multidrug-resistance-associated protein MGr1-Ag is identical to the human 37-kDa laminin receptor precursor. Cell Mol Life Sci. 2002;59:1577–1583.CrossRefGoogle Scholar
  22. 22.
    Marquardt A, Stohr H, White K, Weber BHF. cDNA cloning, genomic structure, and chromosomal location of three members of the human fatty acid desaturase family. Genomics. 2000;66:175–183.CrossRefGoogle Scholar
  23. 23.
    Marzo I, Martinez-Lorenzo MJ, Anel A, et al. Biosynthesis of unsaturated fatty acids in the main cell lineages of human leukemia and lymphoma. Biochim Biophys Acta. 1995;1257:140–148.CrossRefGoogle Scholar

Copyright information

© The Japanese Society of Hematology 2004

Authors and Affiliations

  • Yoshikazu Ichikawa
    • 1
  • Makoto Hirokawa
    • 1
  • Namiko Aiba
    • 1
  • Naohito Fujishima
    • 1
  • Atsushi omatsuda
    • 1
  • Hirobumi Saitoh
    • 1
  • Masaaki Kume
    • 1
  • Ikuo Miura
    • 1
  • Ken-ichi Sawada
    • 2
  1. 1.Department of Internal Medicine IIIAkita University School of MedicineAkitaJapan
  2. 2.Department of Internal MedicineAkita University School of MedicineAkitaJapan

Personalised recommendations