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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1994 Mar;85(3):290–297. doi: 10.1111/j.1349-7006.1994.tb02096.x

Characterization of a Taxol‐resistant Human Small‐cell Lung Cancer Cell Line

Sei Ohta 1,2, Kazuto Nishio 1, Naohiro Kubota 1, Tohru Ohmori 1,2, Yasunori Funayama 1, Tatsuo Ohira 1, Hiroaki Nakajima 2, Mitsuru Adachi 2, Nagahiro Saijo 1,✉
PMCID: PMC5919448  PMID: 7514586

Abstract

Taxol is a novel anticancer agent with activity against a broad range of tumors. It has a unique ability to stabilize polymerized tubulin into microtubule bundles within the cell. We have established a taxol‐resistant human small‐cell lung cancer cell line (H69/Txl) by exposing H69 cells to stepwise increases in taxol concentration. The resistance of H69/Txl cells to taxol was 4.7‐fold that of the original H69 cells: the IC50 values for H69 and H69/Txl were 113.7 ± 56.54 nM and 538.7 ± 214.7 nM by the tetrazolium dye assay, respectively. Removal of the drug from the medium resulted in a 38% decrease in the growth rate of H69/Txl as compared with that in the presence of 30 nM taxol, suggesting that the growth of H69/Txl was partially dependent on taxol. H69/Txl showed higher sensitivity to vinca alkaloids such as vindesine, vincristine and vinblastine than the parental H69. There was no significant difference in intracellular [3H]taxol content between H69 and H69/Txl cells. No MDR‐1 mRNA was detected in H69/Txl by the reverse transcription polymerase chain reaction. There was no significant difference of total and polymerized tubulin content between H69 and H69/Txl cells. Altered mobility of one of the α‐tubulin isoforms in H69/Txl was revealed by using isoelectric focusing and Western blotting with anti‐α‐tubulin antibody. In H69, two α‐tubulin isoforms were observed, whereas three were evident in H69/Txl, two of them comigrating with the isoforms of H69 and the other being more acidic. We observed the increased acetylation of α‐tubulin in H69/Txl cells as compared with that in H69 cells. The acetylation of α‐tubulin may be responsible for the taxol resistance and/or taxol‐dependent growth of H69/Txl.

Keywords: Taxol, Drug resistance, Tubulin, Polymerization, Acetylation

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