Skip to main content
SpringerLink
Log in

Characterization of an etoposide-resistant human small-cell lung cancer cell line

  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Summary

We established an etoposide (VP-16)-resistant human small-cell lung cancer cell line (H69/VP) by stepwise exposure to VP-16. The resistance of H69/VP to VP-16 was 9.4-fold that of the parent cell line (H69/P). H69/VP showed cross-resistance to Adriamycin (ADM), (4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino) carbonyloxy]-1H-pyrano[3′,4′:6,7]indolizino [1,2-b]quinoline-3,14(4H,12H)-dionehydrochloride trihydrate (CPT-11), teniposide (VM-26), vindesine (VDS) and vincristine (VCR). The amount of DNA topoisomerase II (topo.II) was nearly the same in H69/P and H69/VP cells. The catalytic activity of topo.II in H69/VP cells was lower than that in the H69/P line. Accumulation of [3H]-VP-16 in H69/VP was 6.1 – 7.5 times lower than that in H69/P. According to Northern blot analysis, themdr-1 mRNA level in H69/VP was markedly higher than that in H69/P. These findings suggest that H69/VP has a typical multidrug resistance (MDR) phenotype and that alteration of the drug accumulation mediated by P-glycoprotein may play an important role in resistance to VP-16. Reduced topo.II activity may also be associated with VP-16 resistance.

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

Abbreviations

VP-16:

etoposide

ADM:

Adriamycin

CPT-11:

(4S)-4,11-diethyl-4-hydroxy-9-[(4-piperidinopiperidino)carbonyloxy]-1H-pyrano[3′,4′:6,7]indolizino[1, 2-b]quinoline-3,14(4H,12H)-dionehydrochloride trihydrate

VM-26:

teniposide

VDS:

vindesine

VCR:

vincristine

MDR:

multidrug resistance

topo.II:

DNA topoisomerase II

CDDP:

cis-diamminedichloroplatinum(II)

l-PAM:

l-phenylalanine mustard

BLM:

bleomycin

kDNA:

kinetoplast DNA

PBS:

phosphatebuffered saline

SDS:

sodium dodecyl sulfate

EDTA:

ethylendiaminetetraacetic acid

topo.I:

DNA topoisomerase I

CHO:

Chinese hamster ovary

References

  1. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248

    Article  PubMed  CAS  Google Scholar 

  2. Chen GL, Yang L, Rowe TC, Halligan BD, Tewey KM, Liu LF (1984) Nonintercalative antitumor drugs interfere with the breakage-reunion reaction of mammalian DNA topoisomerase II. J Biol Chem 259: 13560

    PubMed  CAS  Google Scholar 

  3. Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156

    Article  PubMed  CAS  Google Scholar 

  4. Deffie AM, Batra JK, Goldenberg GJ (1989) Direct correlation between DNA topoisomerase II activity and cytotoxicity in Adriamycin-sensitive and -resistant P388 leukemia cell lines. Cancer Res 49: 58

    PubMed  CAS  Google Scholar 

  5. Ferguson PJ, Cheng YC (1987) Transient protection of cultured human cells against antitumor agents by 12-O-tetradecanoylphorbol-13-acetate. Cancer Res 47: 433

    PubMed  CAS  Google Scholar 

  6. Ferguson PJ, Fisher MH, Stephenson J, Li DH, Zhou BS, Cheng YC (1988) Combined modalities of resistance in etoposide-resistant human KB cell lines. Cancer Res 48: 5956

    PubMed  CAS  Google Scholar 

  7. Fischer GA, Sartorelli AC (1964) Development, maintenance and assay of drug resistance. Methods Med Res 10: 247

    PubMed  CAS  Google Scholar 

  8. Glisson B, Gupta R, Smallwood-Kentro S, Ross W (1986) Characterization of acquired epipodophyllotoxin resistance in a Chinese hamster ovary cell line: loss of drug-stimulated DNA cleavage activity. Cancer Res 46: 1934

    PubMed  CAS  Google Scholar 

  9. Hill BT, Lock RB (1989) Tumor cell resistance to etoposide (VP-16) — review. In: Kessel D (eds) Resistance to antineoplastic drugs. CRC Press, Boca Raton, Florida, p 185

    Google Scholar 

  10. Hill BT, Whelan RDH, Hosking LK, Shellard SA, Bedford P, Lock RB (1988) Interactions between antitumor drugs and radiation in mammalian tumor cell lines: differential drug responses and mechanisms of resistance following fractionated X-irradiation or continuous drug exposure in vitro. NCI Monogr 6: 177

    PubMed  Google Scholar 

  11. Liu LF, Miller KG (1981) Eukaryotic DNA topoisomerase: two forms of type I DNA topoisomerases from HeLa cell nuclei. Proc Natl Acad Sci USA 78: 3487

    Article  PubMed  CAS  Google Scholar 

  12. Long BH, Musial ST, Brattain MG (1986) DNA breakage in human lung carcinoma cells and nuclei that are naturally sensitive or resistant to etoposide and teniposide. Cancer Res 46: 3809

    PubMed  CAS  Google Scholar 

  13. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265

    PubMed  CAS  Google Scholar 

  14. Marini JC, Miller KG, Englund PE (1980) Decatenation of kinetoplast DNA by topoisomerases. J Biol Chem 255: 4976

    PubMed  CAS  Google Scholar 

  15. Pommier Y, Kerrigan D, Schwartz RE, Swack JA, McCurdy A (1986) Altered DNA topoisomerase II activity in Chinese hamster cells resistant to topoisomerase II inhibitors. Cancer Res 46: 3075

    PubMed  CAS  Google Scholar 

  16. Roninson IB, Chin JE, Choi K, Gros P, Housman DE, Fojo A, Shen DW, Gottesman MM, Pastan I (1986) Isolation of humanmdr-DNA sequences amplified in multidrug-resistant KB carcinoma cells. Proc Natl Acad Sci USA 83: 4538

    Article  PubMed  CAS  Google Scholar 

  17. Rowe T, Kupfer G, Ross W (1985) Inhibition of epipodophyllotoxin cytotoxicity by interference with topoisomerase-mediated DNA cleavage. Biochem Pharmacol 34: 2483

    Article  PubMed  CAS  Google Scholar 

  18. Seeber S, Osieka R, Schmidt CG, Achterrath W, Crooke ST (1982) In vivo resistance towards anthracyclines, etoposide, andcis-diamminedichloroplatinum(II). Cancer Res 42: 4719

    PubMed  CAS  Google Scholar 

  19. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

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

    Koichi Minato, Fumihiko Kanzawa, Kazuto Nishio, Kazuhiko Nakagawa, Yasuhiro Fujiwara & Nagahiro Saijo

  2. First Department of Internal Medicine, Gunma University School of Medicine, 3-39-22 Shouwa-machi, Maebashi, 371, Gunma, Japan

    Koichi Minato

Authors
  1. Koichi Minato
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fumihiko Kanzawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kazuto Nishio
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kazuhiko Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yasuhiro Fujiwara
    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

Rights and permissions

Reprints and permissions

About this article

Cite this article

Minato, K., Kanzawa, F., Nishio, K. et al. Characterization of an etoposide-resistant human small-cell lung cancer cell line. Cancer Chemother Pharmacol 26, 313–317 (1990). https://doi.org/10.1007/BF02897284

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02897284

Keywords

Search

Navigation