Peloruside- and laulimalide-resistant human ovarian carcinoma cells have βI-tubulin mutations and altered expression of βII- and βIII-tubulin isotypes

Arun Kanakkanthara, Anja Wilmes, Aurora O'Brate, Daniel Escuin, Ariane Chan, Ada Gjyrezi, Janet Crawford, Pisana Rawson, Bronwyn Kivell, Peter T. Northcote, Ernest Hamel, Paraskevi Giannakakou, John H. Miller

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35 Scopus citations

Abstract

Peloruside A and laulimalide are potent microtubule-stabilizing natural products with a mechanism of action similar to that of paclitaxel. However, the binding site of peloruside A and laulimalide on tubulin remains poorly understood. Drug resistance in anticancer treatment is a serious problem. We developed peloruside A- and laulimalide-resistant cell lines by selecting 1A9 human ovarian carcinoma cells that were able to grow in the presence of one of these agents. The 1A9-laulimalide resistant cells (L4) were 39-fold resistant to the selecting agent and 39-fold cross-resistant to peloruside A, whereas the 1A9-peloruside A resistant cells (R1) were 6-fold resistant to the selecting agent while they remained sensitive to laulimalide. Neither cell line showed resistance to paclitaxel or other drugs that bind to the taxoid site on b-tubulin nor was there resistance to microtubule-destabilizing drugs. The resistant cells exhibited impaired peloruside A/ laulimalide-induced tubulin polymerization and impaired mitotic arrest. Tubulin mutations were found in the bI-tubulin isotype, R306H or R306C for L4 and A296T for R1 cells. This is the first cell-based evidence to support a b-tubulin-binding site for peloruside A and laulimalide. To determine whether the different resistance phenotypes of the cells were attributable to any other tubulin alterations, the b-tubulin isotype composition of the cells was examined. Increased expression of bII- and bIII-tubulin was observed in L4 cells only. These results provide insight into how alterations in tubulin lead to unique resistance profiles for two drugs, pelorusideAand laulimalide, that have a similar mode of action.

Original languageEnglish (US)
Pages (from-to)1419-1429
Number of pages11
JournalMolecular Cancer Therapeutics
Volume10
Issue number8
DOIs
StatePublished - Aug 1 2011
Externally publishedYes

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ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Kanakkanthara, A., Wilmes, A., O'Brate, A., Escuin, D., Chan, A., Gjyrezi, A., Crawford, J., Rawson, P., Kivell, B., Northcote, P. T., Hamel, E., Giannakakou, P., & Miller, J. H. (2011). Peloruside- and laulimalide-resistant human ovarian carcinoma cells have βI-tubulin mutations and altered expression of βII- and βIII-tubulin isotypes. Molecular Cancer Therapeutics, 10(8), 1419-1429. https://doi.org/10.1158/1535-7163.MCT-10-1057