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Cancer Chemotherapy and Pharmacology
Published in: Cancer Chemotherapy and Pharmacology 2/2012

01-08-2012 | Original Article

DAT-230, a novel microtubule inhibitor, exhibits potent anti-tumor activity by inducing G2/M phase arrest, apoptosis in vitro and perfusion decrease in vivo to HT-1080

Authors: Foxiao Qiao, Daiying Zuo, Xueqi Shen, Huan Qi, Haifeng Wang, Weige Zhang, Yingliang Wu

Published in: Cancer Chemotherapy and Pharmacology | Issue 2/2012

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Abstract

Purpose

The anti-mitotic agent, combretastatin A-4 (CA-4), is the lead compound of a new class of anti-cancer drugs that target tumor vasculature. 2-Methoxy-5-(2-(3, 4, 5-trimethoxyphenyl) thiophen-3-yl) aniline (DAT-230) is a structurally novel CA-4 analog with more stability. We investigated its anti-tumor activity and mechanisms in vitro and in vivo for the first time.

Methods

Cytotoxicity was measured by MTT method. Apoptosis, mitochondria membrane potential (ΔΨm) and NO generation were measured by flow cytometry. Intracellular microtubule network was detected by immunofluorescence experiments. Protein expression was analyzed by Western blotting. In vivo, the anti-tumor activity was assessed using fibrosarcoma xenografts subcutaneously established in BALB/c nude mice. Vasculature perfusion was identified using fluorescent DNA-binding compound Hoechst 33342.

Results

DAT-230 exhibited potent anti-proliferative activity against various cancer cells. DAT-230-treatment in HT-1080 cells resulted in microtubule de-polymerization and G2/M phase arrest preceding apoptosis. Phosphor-cdc2 (thr14/tyr15) reduction, cyclin B1 accumulation and aberrant spindles denoted the cyclin B1-cdc2 complex active and M phase arrest in HT-1080 cells treated with DAT-230. Apoptosis induced by DAT-230 was related with the activation of caspase-9, caspase-3 and PARP cleavage, which were at the downstream of mitochondria. The decrease ratio of Bcl-2/Bax, elevation of NO and disruption of ΔΨm confirmed the causal relationship between DAT-230 and mitochondrial pathway. In vivo, DAT-230 delayed tumor growth, induced tumor perfusion decrease and extensive hemorrhagic-necrosis.

Conclusions

DAT-230 is a promising microtubule inhibitor that has great potential for the treatment of fibrosarcoma in vitro and in vivo. Its potential to be a candidate of anti-cancer agent is worth being further investigated.
Appendix
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Metadata
Title
DAT-230, a novel microtubule inhibitor, exhibits potent anti-tumor activity by inducing G2/M phase arrest, apoptosis in vitro and perfusion decrease in vivo to HT-1080
Authors
Foxiao Qiao
Daiying Zuo
Xueqi Shen
Huan Qi
Haifeng Wang
Weige Zhang
Yingliang Wu
Publication date
01-08-2012
Publisher
Springer-Verlag
Published in
Cancer Chemotherapy and Pharmacology / Issue 2/2012
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI
https://doi.org/10.1007/s00280-012-1907-x

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