Proapoptotic activity of Ukrain is based on Chelidonium majusL. alkaloids and mediated via a mitochondrial death pathwayReport as inadecuate




Proapoptotic activity of Ukrain is based on Chelidonium majusL. alkaloids and mediated via a mitochondrial death pathway - Download this document for free, or read online. Document in PDF available to download.

BMC Cancer

, 6:14

First Online: 17 January 2006Received: 13 September 2005Accepted: 17 January 2006DOI: 10.1186-1471-2407-6-14

Cite this article as: Habermehl, D., Kammerer, B., Handrick, R. et al. BMC Cancer 2006 6: 14. doi:10.1186-1471-2407-6-14

Abstract

BackgroundThe anticancer drug Ukrain NSC-631570 which has been specified by the manufacturer as semisynthetic derivative of the Chelidonium majus L. alkaloid chelidonine and the alkylans thiotepa was reported to exert selective cytotoxic effects on human tumour cell lines in vitro. Few clinical trials suggest beneficial effects in the treatment of human cancer. Aim of the present study was to elucidate the importance of apoptosis induction for the antineoplastic activity of Ukrain, to define the molecular mechanism of its cytotoxic effects and to identify its active constituents by mass spectrometry.

MethodsApoptosis induction was analysed in a Jurkat T-lymphoma cell model by fluorescence microscopy chromatin condensation and nuclear fragmentation, flow cytometry cellular shrinkage, depolarisation of the mitochondrial membrane potential, caspase-activation and Western blot analysis caspase-activation. Composition of Ukrain was analysed by mass spectrometry and LC-MS coupling.

ResultsUkrain turned out to be a potent inducer of apoptosis. Mechanistic analyses revealed that Ukrain induced depolarisation of the mitochondrial membrane potential and activation of caspases. Lack of caspase-8, expression of cFLIP-L and resistance to death receptor ligand-induced apoptosis failed to inhibit Ukrain-induced apoptosis while lack of FADD caused a delay but not abrogation of Ukrain-induced apoptosis pointing to a death receptor independent signalling pathway. In contrast, the broad spectrum caspase-inhibitor zVAD-fmk blocked Ukrain-induced cell death. Moreover, over-expression of Bcl-2 or Bcl-xL and expression of dominant negative caspase-9 partially reduced Ukrain-induced apoptosis pointing to Bcl-2 controlled mitochondrial signalling events.

However, mass spectrometric analysis of Ukrain failed to detect the suggested trimeric chelidonine thiophosphortriamide or putative dimeric or monomeric chelidonine thiophosphortriamide intermediates from chemical synthesis. Instead, the Chelidonium majus L. alkaloids chelidonine, sanguinarine, chelerythrine, protopine and allocryptopine were identified as major components of Ukrain.

Apart from sanguinarine and chelerythrine, chelidonine turned out to be a potent inducer of apoptosis triggering cell death at concentrations of 0.001 mM, while protopine and allocryptopine were less effective. Similar to Ukrain, apoptosis signalling of chelidonine involved Bcl-2 controlled mitochondrial alterations and caspase-activation.

ConclusionThe potent proapoptotic effects of Ukrain are not due to the suggested -Ukrain-molecule- but to the cytotoxic efficacy of Chelidonium majus L. alkaloids including chelidonine.

List of abbreviationscFLIPcellular FADD-like interleukin-1 converting enzyme FLICE inhibitory protein

FADDFas-associated death domain

PARPpolyADP-ribosepolymerase

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2407-6-14 contains supplementary material, which is available to authorized users.

Daniel Habermehl, Bernd Kammerer contributed equally to this work.

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Author: Daniel Habermehl - Bernd Kammerer - René Handrick - Therese Eldh - Charlotte Gruber - Nils Cordes - Peter T Daniel - Ludw

Source: https://link.springer.com/







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