Microtubule Alterations and Mutations Induced by Desoxyepothilone B

Verrills, Nicole M.; Flemming, Claudia L.; Liu, Marjorie; Ivery, Michael T. G.; Cobon, Gary S.; Norris, Murray D.; Haber, Michelle; Kavallaris, Maria

doi:10.1016/s1074-5521(03)00141-8

Cited by 107 publications

(95 citation statements)

References 44 publications

(1 reference statement)

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“…Growth inhibition assays were done as previously described (22). Briefly, cells were seeded at 3,750 per well (HMEC-1 and MRC-5) or 1,500 per well (BMH29L) in 96-well gelatin-coated plates.…”

Section: Methodsmentioning

confidence: 99%

ENMD-1198, a New Analogue of 2-Methoxyestradiol, Displays Both Antiangiogenic and Vascular-Disrupting Properties

Pasquier

Sinnappan

Munoz

et al. 2010

Molecular Cancer Therapeutics

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The formation of a new vascular network by angiogenesis is a key driver in tumor growth and metastasis, making this an attractive therapeutic target. Different strategies are being developed to either prevent tumor angiogenesis or disrupt the tumor vasculature already in place. In this in vitro study, we investigated the antivascular properties of ENMD-1198, a new anticancer drug currently in clinical trials. ENMD-1198 is a new analogue of 2-methoxyestradiol, a microtubule-targeting agent that has shown promising results in the treatment of multiple myeloma and hormone-refractory prostate cancer. Using both bone marrowderived and dermal microvascular endothelial cell lines, we analyzed the effect of ENMD-1198 on the different functions of endothelial cells involved in angiogenesis. In both cell lines, ENMD-1198 was more potent than 2-methoxyestradiol at inhibiting endothelial cell proliferation, motility, migration, and morphogenesis. In addition, ENMD-1198 induced a significant decrease in vascular endothelial growth factor receptor-2 protein expression in endothelial cells. Furthermore, videomicroscopy experiments showed that ENMD-1198 was able to completely disrupt preformed vascular structures within 2 hours. This vascular-disrupting activity was associated with extensive depolymerization of the microtubule network and accumulation of actin stress fibers and large focal adhesions in vascular endothelial cells. Collectively, our results show that this new compound displays potent antivascular properties, and this study provides important insights into the mechanism of action of this promising new anticancer drug.

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Section: Methodsmentioning

confidence: 99%

ENMD-1198, a New Analogue of 2-Methoxyestradiol, Displays Both Antiangiogenic and Vascular-Disrupting Properties

Pasquier

Sinnappan

Munoz

et al. 2010

Molecular Cancer Therapeutics

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“…Two-Dimensional PAGE and Immunoblotting Analysis Two-dimensional PAGE of parental CCRF-CEM and 2ME2-resistant cells was carried out as described previously (21), with minor modifications. Isolated microtubule pellets from CCRF-CEM and 2ME2-resistant cells were solubilized with isoelectric focusing buffer (7 mol/L urea, 2 mol/L thiourea, 2% CHAPS, 2% sulfobetaines 3-10, 1% amidosulfobetaine-14, 2 mol/L tributylphosphine, 65 mmol/L DTT, 1% carrier ampholytes 3-10, 1% carrier ampholytes 4-6, 0.01% bromophenol blue).…”

Section: Isolation Of Tubulin From Cell Linesmentioning

confidence: 99%

Class I β-tubulin mutations in 2-methoxyestradiol-resistant acute lymphoblastic leukemia cells: implications for drug-target interactions

Liaw

Salam

McKay

et al. 2008

Molecular Cancer Therapeutics

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2-Methoxyestradiol (2ME2) is a naturally occurring derivative of estradiol that has been shown to be an active small molecule that has antitumor and antiangiogenic properties. 2ME2 binds to B-tubulin near the colchicinebinding site, inhibits microtubule polymerization, and induces mitotic arrest. To improve understanding of the mechanisms of action and resistance to 2ME2, we selected leukemia cells, CCRF-CEM, that display increasing resistance to 2ME2, and three of the highly resistant sublines were chosen for detailed analysis. The 2ME2 cells selected in 7.2 to 28.8 Mmol/L were found to be 47-to 107-fold resistant to 2ME2 and exhibited low levels of cross-resistance to vinblastine. Two of the lowest 2ME2-resistant sublines were significantly hypersensitive to colchicine and epothilone B, but the hypersensitive effects were lost in the highest 2ME2-resistant subline. Moreover, 2ME2-resistant cells require 10-fold higher concentrations of 2ME2 to induce G 2 -M cell cycle arrest and have higher amounts of tubulin polymer compared with parental cells. Gene and protein sequencing revealed four class I B-tubulin mutations, S25N, D197N, A248T, and K350N, in the 2ME2-resistant cells. The S25N mutation is within the paclitaxel-binding site, whereas A248T and K350N are within the colchicine-binding site on B-tubulin, yet the resistant cells were not cross-resistant to paclitaxel or colchicine. This strongly suggests that the mutations have induced conformational changes to the binding site that resulted in 2ME2 resistance. The 2ME2-resistant leukemia cells provide novel insights into microtubule stability and drug-target interactions. [Mol Cancer Ther 2008;7(10):3150 -9]

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“…5 Furthermore, b-tubulin mutations in these regions have been suggested to be related with tumor cell chemosensitivity to microtubule-binding drugs. 4,6 Paclitaxel is an antineoplastic drug extensively used in breast cancer treatment binding the b-tubulin subunits on the microtubule, blocking cells in G 2 /M phase and leading to apoptotic cell death. 7 These cytotoxic effects have been reported to be clearly related with the induction of microtubule assembly and promotion of microtubule bundle formation.…”

mentioning

confidence: 99%

Cytoskeleton and paclitaxel sensitivity in breast cancer: The role of β‐tubulins

Tommasi¹,

Mangia²,

Lacalamita³

et al. 2007

Intl Journal of Cancer

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139

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The antineoplastic effect of paclitaxel is mainly related to its ability to bind the b subunit of tubulin, thus preventing tubulin chain depolarization and inducing apoptosis. The relevance of the Class I b-tubulin characteristics have also been confirmed in the clinical setting where mutations of paclitaxel-binding site of b-tubulin Class I have been related to paclitaxel resistance in non small cell lung and ovarian cancers. In the present study, we verified the hypothesis of a relationship between molecular alterations of b-tubulin Class I and paclitaxel sensitivity in a panel of breast cell lines with different drug IC 50 . The Class I b-tubulin gene cDNA has been sequenced detecting heterozygous missense mutations (exon 1 and 4) only in MCF-7 and SK-BR-3 lines. Furthermore, the expression (at both mRNA and protein level) of the different isotypes have been analyzed demonstrating an association between low cell sensitivity to paclitaxel and Class III b-tubulin expression increasing. Antisense oligonucleotide (ODN) experiments confirmed that the inhibition of Class III b-tubulin could at least partially increase paclitaxel-chemosensitivity. The hypothesis of a relationship between b-tubulin tumor expression and paclitaxel clinical response has been finally verified in a series of 92 advanced breast cancer patients treated with a first line paclitaxel-based chemotherapy. Thirty-five percent (95% CI: 45-31) of patients with high Class III b-tubulin expression showed a disease progression vs. only 7% of patients with low expression (35% vs. 7%, p < 0.002). Our study suggests that Class III b-tubulin tumor expression could be considered a predictive biomarker of paclitaxel-clinical resistance for breast cancer patients. ' 2007 Wiley-Liss, Inc.

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Microtubule Alterations and Mutations Induced by Desoxyepothilone B

Cited by 107 publications

References 44 publications

ENMD-1198, a New Analogue of 2-Methoxyestradiol, Displays Both Antiangiogenic and Vascular-Disrupting Properties

ENMD-1198, a New Analogue of 2-Methoxyestradiol, Displays Both Antiangiogenic and Vascular-Disrupting Properties

Class I β-tubulin mutations in 2-methoxyestradiol-resistant acute lymphoblastic leukemia cells: implications for drug-target interactions

Cytoskeleton and paclitaxel sensitivity in breast cancer: The role of β‐tubulins

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