Skip to main content
SpringerLink
Log in

Apoptosis induced by etoposide in small-cell lung cancer cell lines

  • Original Articles
  • DNA Fragmentation, Apoptosis, Lung Cancer Cell
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

The DNA fragmentation, a parameter of apoptosis, in non-small (NSCLC) and small (SCLC) cell lung cancer cell lines (N231 and PC-9) was evaluated. The DNA fragmentation in SCLC lines, but not in NSCLC lines, was observed in overgrown cells without exposure to anticancer drugs. In etoposide (VP-16)-treated N231 but not PC-9 cells, DNA fragmentation continued to increase up to 42 h, and the increase was dependent on the concentration of VP-16. The endonuclease activity of VP-16-treated N231, but not PC-9, cells required both Ca2+ and Mg2+ for full activity. It was elevated in a time- and concentration-dependent manner. As this activity was not affected by addition of cycloheximide, the activation of the endonuclease activity without protein synthesis may be involved in VP-16-induced cytotoxicity in N231.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Kerr, JFR, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26: 239–257

    Google Scholar 

  2. Wyllie AH, Kerr JFR, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68: 251–306

    Google Scholar 

  3. Cohen JJ (1993) Apoptosis. Immunol Today 14: 126–130

    Google Scholar 

  4. Duvall E, Wyllie AH (1986) Death and the cell Immunol Today 7: 115–119

    Google Scholar 

  5. Barry MA, Behnke CA, Eastman A (1990) Activation of programmed cell death (apoptosis) by cisplatin, other anticancer drugs, toxins and hyperthermia. Biochem Pharmacol 40: 2353–2362

    Google Scholar 

  6. Stephens LC, Ang KK, Schultheiss TE, Milas L, Meyn RE (1991) Apoptosis in irradiated murine tumors. Radiat Res 127: 308–316

    Google Scholar 

  7. Takano YS, Harmon BV, Kerr JF (1991) Apoptosis induced by mild hyperthermia in human and murine tumour cell lines: a study using electron microscopy and DNA gel electrophoresis. J Pathol 163: 329–336

    Google Scholar 

  8. Wylle AH (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogeneous endonuclease activation. Nature 284: 555

    Google Scholar 

  9. Nieto MA, Gonzalez A, Gambon F, Diaz EF, Lopez RA (1992) Apoptosis in human thymocytes after treatment with glucocorticoids. Clin Exp Immunol 88: 341–344

    Google Scholar 

  10. Morimoto H, Bonavida B (1992) Diphtheria toxin- andPseudomonas A toxin-mediated apoptosis. ADP ribosylation of elongation factor-2 is required for DNA fragmentation and cell lysis and synergy with tumor necrosis factor-alpha. J Immunol 149: 2089–2094

    Google Scholar 

  11. Larrick JW, Wright SC (1990) Cytotoxic mechanism of tumor necrosis factor-alpha. Faseb J 4: 3215–3223

    Google Scholar 

  12. Obeid LM, Linardic CM, Karolak LA, Hannun YA (1993) Programmed cell death induced by ceramide. Science 259: 1769–1771

    Google Scholar 

  13. Curtin NJ, Harris AL, Aherne GW (1991) Mechanism of cell death following thymidylate synthase inhibition: 2′-deoxyuridine-5′-triphosphate accumulation, DNA damage, and growth inhibition following exposure to CB3717 and dipyridamole. Cancer Res 51: 2346–2352

    Google Scholar 

  14. Li JC, Kaminskas E (1987) Progressive formation of DNA lesions in cultured Ehrlich ascites tumor cells treated with hydroxyurea. Cancer Res 47: 2755–2758

    Google Scholar 

  15. Ling YH, Priebe W, Perez SR (1993) Apoptosis induced by anthracycline antibiotics in P388 parent and multidrug-resistant cells. Cancer Res 53: 1845–1852

    Google Scholar 

  16. Bertrand R, Sarang M, Jenkin J, Kerrigan D, Pommier Y (1991) Differential induction of secondary DNA fragmentation by topoisomerase II inhibitors in human tumor cell lines with amplified c-myc expression. Cancer Res 51: 6280–6285

    Google Scholar 

  17. O'Connor PM, Wassermann K, Sarang M, Magrath I, Bohr VA, Kohn KW (1991) Relationship between DNA cross-links, cell cycle, and apoptosis in Burkitt's lymphoma cell lines differing in sensitivity to nitrogen mustard. Cancer Res 51: 6550–6557

    Google Scholar 

  18. Ohmori T, Podack ER, Nishio K, Takahashi M, Miyahara Y, Takeda Y, Kubota N, Funayama Y, Ogasawara H, Ohira T, Ohta S, Saijo N (1993) Apoptosis of lung cancer cells caused by some anti-cancer agents (MMC, CPT-11, ADM) is inhibited bybcl-2. Biochem Biophys Res Commun 192: 30–36

    Google Scholar 

  19. Bhalla K, Ibrado AM, Tourkina E, Tang C, Mahoney ME, Huang Y (1993) Taxol induces internucleosomal DNA fragmentation associated with programmed cell death in human myeloid leukemia cells. Leukemia 7: 563–568

    Google Scholar 

  20. Eastman A (1990) Activation of programmed cell death by anticancer agents: cisplatin as a model system. Cancer Cell 2: 275–280

    Google Scholar 

  21. McConkey DJ, Hartzell P, Nicotera P, Orrenius S (1989) Calcium-activated DNA fragmentation kills immature thymocytes. Faseb J 3: 1843–1849

    Google Scholar 

  22. Mosmann T (1983) Rapid coorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity. J Immunol Meth 65: 55–63

    Google Scholar 

  23. Smith CA, Williams GT, Kingston R, Jenkinson EJ, Owen JJT (1989) Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature 337: 181–184

    Google Scholar 

  24. Lee KW, Bindereif A, Green MR (1988) A small-scale procedure for preparation of nuclear extracts that support efficient transcription and pre-mRNA splicing. Gene Anal Techn 5: 22–31

    Google Scholar 

  25. Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of proteins utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Google Scholar 

  26. Hashida T, Tanaka Y, Matsunami N, Yoshihara K, Kamiya T, Tanigawa Y, Koide S (1982) Purification and properties of bull seminal plasma Ca2+, Mg2+-dependent endonuclease. J Biol Chem 257: 13114–13119

    Google Scholar 

  27. Giannakis C, Forbes IJ, Zalewski PD (1991) Ca2+/Mg2+-dependent nuclease: tissue distribution, relationship to inter-nucleosomal DNA fragmentation and inhibition by Zn2+ Biochem Biophys Res Commun 181: 915–920

    Google Scholar 

  28. Walker PR, Smith C, Youdale T, Leblanc J, Whitfield JF, Sikorska M (1991) Topoisomerase II-reactive chemotherapeutic drugs induce apoptosis in thymocytes. Cancer Res 51: 1078–1085

    Google Scholar 

  29. Shimizu T, Kubota M, Tanizawa A, Sano H, Kasai Y, Hashimoto H, Akiyama Y, Mikawa H (1990) Inhibition of both etoposide-induced DNA fragmentation and activation of poly(ADP-ribose) synthesis by zinc ion. Biochem Biophys Res Commun 169: 1172–1177

    Google Scholar 

  30. Roy C, Brown DL, Little JE, Valentine BK, Walker PR, Sikorska M, Leblanc J, Chaly N (1992) The topoisomerase II inhibitor teniposide (VM-26) induces apoptosis in unstimulated mature murine lymphocytes. Exp Cell Res 200: 416–424

    Google Scholar 

  31. Rice WG, Hillyer CD, Harten B, Schaeffer CA, Dorminy M, Lackey D3, Kirsten E, Mendeleyev J, Buki KG, Hakam A, Kun E (1992) Induction of endonuclease-mediated apoptosis in tumor cells by C-nitroso-substituted ligands of poly(ADP-ribose) polymerase. Proc Natl Acad Sci USA 89: 7703–7707

    Google Scholar 

  32. Kubo S, Matsutani M, Nakagawa K, Ogura T, Esumi H, Saijo N (1992) Participation of poly(ADP-ribose) polymerase in the drug sensitivity in human lung cancer cell lines. J Cancer Res Clin Oncol 118: 244–248

    Google Scholar 

  33. Kaufmann SH (1989) Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic anticancer drugs: a cautionary note. Cancer Res 49: 5870–5878

    Google Scholar 

  34. Rubin E, Kharbanda S, Gunji H, Kufe D (1991) Activation of the c-jun protooncogene in human myeloid leukemia cells treated with etoposide. Mol Pharmacol 39: 697–701

    Google Scholar 

  35. Lock RB, Ross WE (1990) Possible role for p34cdc2 kinase in etoposide-induced cell death of Chinese hamster ovary cells. Cancer Res 50: 3767–3771

    Google Scholar 

  36. Lock RB, Ross WE (1990) Inhibition of p34cdc2 kinase activity by etoposide or irradiation as a mechanism of G2 arrest in Chinese hamster ovary cells. Cancer Res 50: 3761–3766

    Google Scholar 

  37. Evan GI, Wyllie AH, Gilbert CS, Littlewood TD, Land H, Brooks M, Walters CM, Penn LZ, Hancock DC (1992) Induction of apoptosis in fibroblasts byc-myc protein. Cell 69: 119–128

    Google Scholar 

  38. Dive C, Hickman JA (1991) Drug-target interactions: only the first step in the commitment to a programmed cell death? Br J Cancer 64: 192–196

    Google Scholar 

  39. Bissonnette RP, Echeverri F, Mahboubi A, Green DR (1992) Apoptotic cell death induced byc-myc is inhibited bybcl-2. Nature 359: 552–554

    Google Scholar 

  40. Fanidi A, Harrington EA, Evan GI (1992) Cooperative interaction betweenc-myc andbcl-2 proto-oncogenes. Nature 359: 554–556

    Google Scholar 

  41. Hfartwell LH, weinert TA (1989) Checkpoints: controls that ensure the order of cell cycle events. Science 246: 629–633

    Google Scholar 

  42. Sorenson CM, Eastman A (1988) Influence ofcis-diammine digression in excision repair proficinent and deficient Chinese hamster ovary cells. Cancer Res 48: 6703–6707

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Pharmacology Division, National Cancer Center Research Institute, Tsukiji 5-1-1, Chuo-ku, 104, Tokyo, Japan

    Sachiyo Okamoto-Kubo, Kazuto Nishio, Yuji Heike, Mitsuji Yoshida, Tohru Ohmori & Nagahiro Saijo (Chief)

Authors
  1. Sachiyo Okamoto-Kubo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kazuto Nishio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuji Heike
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mitsuji Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tohru Ohmori
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Nagahiro Saijo
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported, in part, by grants-in-Aid for Cancer Research from the Ministry of Health and Welfare, the Comprehensive Ten-year Strategy for Cancer Control and the Ministry of Education, Science and Culture, Support from the Bristol Myers Squibb Foundation is also appreciated

Rights and permissions

Reprints and permissions

About this article

Cite this article

Okamoto-Kubo, S., Nishio, K., Heike, Y. et al. Apoptosis induced by etoposide in small-cell lung cancer cell lines. Cancer Chemother. Pharmacol. 33, 385–390 (1994). https://doi.org/10.1007/BF00686267

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00686267

Keywords

Search

Navigation