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The effect of DB-67, a lipophilic camptothecin derivative, on topoisomerase I levels in non-small-cell lung cancer cells

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Abstract

Purpose

To determine the in vitro drug sensitivity of two non-small-cell lung cancer cell lines after treatment with the novel lipophilic camptothecin derivative, 7-tert-butyldimethylsilyl-10-hydroxycamptothecin (DB-67), to determine if topoisomerase I protein levels decrease after treatment with DB-67, and to assess the duration and extent of topoisomerase I modulation after DB-67 exposure, in order to provide information about drug resistance that may be useful in determining an appropriate dosing schedule for DB-67.

Methods

The growth inhibition of the non-small-cell lung cancer cell lines A549 and H460 after exposure to DB-67 was evaluated with the MTS assay. A549 and H460 cells were treated for various times with DB-67 and topoisomerase I levels were determined by western blot analysis. In addition, A549 and H460 cells were treated with DB-67 for 24 h and topoisomerase I levels were determined by western blot analysis daily for 1 week after drug removal.

Results

DB-67 inhibited the growth of both A549 and H460 cells grown in culture; the A549 cells were more resistant to the cytotoxic effects of DB-67 than H460 cells. Notably, A549 cells had approximately one-half the baseline topoisomerase I than H460 cells. Topoisomerase I protein levels significantly decreased after 8–18 h of exposure to DB-67. Both A549 and H460 cells treated with DB-67 for 24 h had only negligible amounts of topoisomerase I at the end of treatment. However, within 24 h of drug removal topoisomerase I levels returned to near baseline levels in both cell lines.

Conclusions

The decrease in topoisomerase I levels caused by DB-67 may represent a mechanism of resistance to this novel camptothecin derivative. Dosing DB-67 once every 48–72 h may maximize the interaction of the drug with topoisomerase I and should be considered as a potential dosing schedule in the preclinical and clinical development of this compound.

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Acknowledgements

The authors are grateful to the University of Kentucky Research and Graduate Studies for their fellowship support of A.K. Bence.

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Authors and Affiliations

  1. Division Pharmacy Practice and Science, 202 College of Pharmacy, University of Kentucky, Lexington, KY 40536-0082, USA

    Aimee K. Bence, Cynthia A. Mattingly & Val R. Adams

  2. Tigen Pharmaceutical, Inc., 235 Bolivar Street, Lexington, KY 40508, USA

    Thomas G. Burke

  3. Division of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536-0082, USA

    Thomas G. Burke

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  1. Aimee K. Bence
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  2. Cynthia A. Mattingly
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  4. Val R. Adams
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Correspondence to Val R. Adams.

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Bence, A.K., Mattingly, C.A., Burke, T.G. et al. The effect of DB-67, a lipophilic camptothecin derivative, on topoisomerase I levels in non-small-cell lung cancer cells. Cancer Chemother Pharmacol 54, 354–360 (2004). https://doi.org/10.1007/s00280-004-0804-3

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