Tumor Biology

, Volume 32, Issue 2, pp 399–408 | Cite as

Establishment of three cisplatin-resistant endometrial cancer cell lines using two methods of cisplatin exposure

  • Yasukazu Sagawa
  • Atsuya Fujitoh
  • Hirotaka Nishi
  • Hiroe Ito
  • Tamaki Yudate
  • Keiichi Isaka
Research Article

Abstract

Using the endometrial cancer cell line EI established in our department, we attempted to establish cisplatin (CDDP)-resistant cell lines by incremental exposure and high concentration exposure methods. Three CDDP-resistant cell lines were isolated, which could be distinguished by morphological differences.
  1. 1.

    Upon acquiring CDDP resistance, the cells tended to become small and grow in a floating state. This tendency was especially marked when using incremental exposure method. Using the incremental exposure method, a cell line obtained by isolating and culturing only adherent cells was designated EICR-Ia, and a cell line established by culturing only floating cells was designated EICR-If. A cell line obtained by the high concentration exposure method was designated EICR-II.

     
  2. 2.

    Upon acquiring CDDP resistance, tumor markers such as TPA and LDH increased, while proliferative capability of the cells was lowered.

     
  3. 3.

    The invasion capability was diminished in EICR-If cells, but was increased in EICR-Ia and EICR-II cells.

     
  4. 4.

    Following exposure to CDDP, the intracellular platinum concentrations were markedly elevated in EI and EICR-If cells, whereas the increase was mild in EICR-Ia and EICR-II cells and the concentration was lower than that in parent EI cells.

     
  5. 5.

    Studies of drug resistance gene expression revealed increased expression of MDR1, GSTπ, and Topo-II in EICR-If cells; increased expression of GSTπ in EICR-II cells; but no expression of any of the genes in EICR-Ia cells.

     
  6. 6.

    Analyses of cancer- and apoptosis-related genes showed increased expressions of Bcl-2, c-Myc, p53, and ICE in EICR-If cells.

     
  7. 7.

    Upon acquiring CDDP resistance, sensitivity to mitomycin and adriamycin decreased, but sensitivity to etoposide and 5-fluorouracil increased.

     

The findings indicate that the mechanisms of CDDP resistance are different in the three cell lines.

Keywords

Endometrial cancer Cisplatin (CDDP) Multidrug resistance 

References

  1. 1.
    Tominaga S et al. Cancer mortality and morbidity statistics. Tokyo: Japan Scientific Societies; 1994.Google Scholar
  2. 2.
    Rosenberg B, VanCamp L, Krigas T. Inhibition of cell division in escherichia coli by electrolysis products from a platinum electrode. Nature. 1965;205:698–9.CrossRefPubMedGoogle Scholar
  3. 3.
    Muss HB. Chemotherapy of metastatic endometrial cancer. Semin Oncol. 1994;21:107–13.PubMedGoogle Scholar
  4. 4.
    Thigpen JT, Blessing JA, Lagasse LD, DiSaia PJ, Homesley HD. Phase II trial of cisplatin as secondline chemotherapy in patients with advanced or recurrent endometrial carcinoma. A gynecologic oncology group study. Am J Clin Oncol. 1984;7:253–6.CrossRefPubMedGoogle Scholar
  5. 5.
    Deppe G, Cohen CJ, Bruckner HW. Treatment of advanced endometrial adenocarcinoma with cis-dichlorodiammine platinum (II) after intensive prior therapy. Gynecol Oncol. 1980;10:51–4.CrossRefPubMedGoogle Scholar
  6. 6.
    Seski JC, Edwards CL, Herson J, Rutledge FN. Cisplatin chemotherapy for disseminated endometrial cancer. Obstet Gynecol. 1982;59:225–8.PubMedGoogle Scholar
  7. 7.
    Burke TW, Stringer CA, Morris M, Freedman RS, Gershenson DM, Kavanagh JJ, et al. Prospective treatment of advanced or recurrent endometrial carcinoma with cisplatin, doxorubicin, and cyclophosphamide. Gynecol Oncol. 1991;40:264–7.CrossRefPubMedGoogle Scholar
  8. 8.
    Barrete RJ, Blessing JA, Homesley HD, Twiggs L, Webster KD. Circadian-timed combination doxorubicin-cisplatin chemotherapy for advanced endometrial carcinoma. Am J Clin Oncol. 1993;16:494–6.CrossRefGoogle Scholar
  9. 9.
    Belliveau JF, Posner MR, Ferrari L, Crabtree GW, Cummings FJ, Wiemann MC, et al. Cisplatin administered as a continuous 5-day infusion: plasma platinum levels and urine platinum excretion. Cancer Treat Rep. 1986;70:1215–7.PubMedGoogle Scholar
  10. 10.
    Yudate T, Isaka K, Okabe K, Takayama M. Establishment and characterization of the new cell line (EI) from a human endometrial adenocarcinoma. (in Japanese with English abstract) HUMAN CELL. 1995;8(1):43–7.PubMedGoogle Scholar
  11. 11.
    Juliano RL, Ling V. A surface glycoprotein modulating drug permeability in Chinease hamster ovary cell mutants. Biochim Biophys Acta. 1976;455:152–62.CrossRefPubMedGoogle Scholar
  12. 12.
    Ueda K, Clark DP, Chen CJ, Roninson IB, Gottesman MM, Pastan I. The human multidrug resistance (mdr1) gene. J Biol Chem. 1987;262:505–8.PubMedGoogle Scholar
  13. 13.
    Ueda K, Cardarelli C, Gottesman MM, Pastan I. Expression of full-length cDNA for the human “MDR1” gene confers resistance to colchicine, doxorubicin, and vinblastine. Proc Natl Acad Sci USA. 1987;84:3004–8.CrossRefPubMedGoogle Scholar
  14. 14.
    Gottesman MM, Pastan I. The multidrug transporter, a double-edged sword. J Biol Chem. 1988;263:12163–6.PubMedGoogle Scholar
  15. 15.
    Bradley G, Juranka PF, Ling V. Mechanism of multidrug resistance. Biochim Biophys Acta. 1998;948:87–128.Google Scholar
  16. 16.
    Goldstein LJ, Galski H, Fojo A, Willingham M, Lai SL, Gazdar A, et al. Expression of a multidrug resistance gene in human cancer. J Natl Cancer Inst. 1989;81:116–24.CrossRefPubMedGoogle Scholar
  17. 17.
    Cole SP, Deeley RG. Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science. 1992;258:1650–4.CrossRefPubMedGoogle Scholar
  18. 18.
    Loe DW, Deeley RG, Cole SP. Biology of the multidrug resistance protein, MRP. Eur J Cancer. 1996;32:945–57.CrossRefGoogle Scholar
  19. 19.
    Godwin AK, Meister A, O’Dwyer PJ, Huang CS, Hamilton TC, Anderson ME. High resistance to cisplatin in human ovarian cancer cell lines is associated with marked increase of glutathione synthesis. Proc Natl Acad Sci USA. 1992;89:3070–4.CrossRefPubMedGoogle Scholar
  20. 20.
    Lai GM, Moscow JA, Alvarez MJ, Fojo AT, Bates SE. Contribution of glutathione and glutathione-dependent enzymes in the reversal of adriamycin resistance in colon carcinoma cell lines. Int J Cancer. 1991;49:688–95.CrossRefPubMedGoogle Scholar
  21. 21.
    Xu BH, Gupta V, Singh SH. Mitomycin C sensitivity in human bladder cancer cells: possible role of glutathione and glutathione transferase in resistance. Arch Biochem Biophys. 1994;308:164–70.CrossRefPubMedGoogle Scholar
  22. 22.
    Sriram R, Ali-Osman F, Livingston R, et al. Modulation of topoisomerase (Topo) II on the kinetics of formation and repair of cis-platinum (cis-DDP) in human tumor cells sensitive and resistant to cis-DDP. Proc Am Assoc Cancer Res. 1990;31:335.Google Scholar
  23. 23.
    Dejong S, Zijlstra JG, Mulder MH, Devries EG. Lack of cross-resistance to fustriecin in a human small-cell lung carcinoma cell line showing topoisomerase II-related drug resistance. Cancer Chemother Pharmacol. 1991;28:461–4.CrossRefGoogle Scholar
  24. 24.
    Waud WR, Harrison SDJ, Gilbert KS, Laster Jr WR, Griswold Jr DP. Antitumor drug cross-resistance in vivo in a cisplatin-resistant murine P388 leukemia. Cancer Chemother Pharmacol. 1991;27:456–63.CrossRefPubMedGoogle Scholar
  25. 25.
    Liu LF, Rowe TC, Yang L, Tewey KM, Chen GL. Cleavage of DNA by mammalian DNA topoisomerase II. J Biol Chem. 1983;258:15365–70.PubMedGoogle Scholar
  26. 26.
    Hannun YA. Apoptosis and the dilemma of cancer chemotherapy. Blood. 1997;89:1845–53.PubMedGoogle Scholar
  27. 27.
    Glinsky GV, Glinsky VV, Ivanova AB, Hueser CJ. Apoptosis and metastasis: increased resistance of metastatic cancer cells is associated with the profound deficiency of apoptosis execution mechanisms. Cancer Lett. 1997;115:185–93.CrossRefPubMedGoogle Scholar
  28. 28.
    Oltvai ZN, Korsmeyer SJ. Checkpoints of dueling dimers foil death wishes. Cell. 1994;79:189–92.CrossRefPubMedGoogle Scholar
  29. 29.
    Knudson CM, Korsmeyer SJ. Bcl-2 and Bax function independently to regulate cell death. Nat Genet. 1997;16:358–63.CrossRefPubMedGoogle Scholar
  30. 30.
    Thornberry NA, Bull HG, Calaycay JR, Chapman KT, Howard AD, Kostura MJ, et al. A novel heterodimeric cysteine protease is required for interleukin-1βprocessing in monocytes. Nature. 1992;356:768–74.CrossRefPubMedGoogle Scholar
  31. 31.
    Lowe SW, Bodis S, MaClatchey A, Remington L, Ruley HE, Fisher DE, et al. p53 status and the efficacy of cancer therapy in vivo. Science. 1994;266:807–10.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2010

Authors and Affiliations

  • Yasukazu Sagawa
    • 1
  • Atsuya Fujitoh
    • 1
  • Hirotaka Nishi
    • 1
  • Hiroe Ito
    • 1
  • Tamaki Yudate
    • 1
  • Keiichi Isaka
    • 1
  1. 1.Department of Obstetrics and GynecologyTokyo Medical UniversityTokyoJapan

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