Helenalin bypasses Bcl-2-mediated cell death resistance by inhibiting NF-κB and promoting reactive oxygen species generation.

Evasion of cell death by overexpression of anti-apoptotic proteins, such as Bcl-2, is commonly observed in cancer cells leading to a lack of response to chemotherapy. Hence, there is a need to find new chemotherapeutic agents that are able to overcome chemoresistance mediated by Bcl-2 and to understand their mechanisms of action. Helenalin, a sesquiterpene lactone (STL), induces cell death and abrogates clonal survival in a highly apoptosis-resistant Bcl-2 overexpressing Jurkat cell line as well as in two other Bcl-2 overexpressing solid tumor cell lines (mammary MCF-7; pancreatic L6.3pl). This effect is not achieved by directly affecting the mitochondria-protective function of Bcl-2 in the intrinsic pathway of apoptosis since Bcl-2 overexpressing Jurkat cells do not show cytochrome c release and dissipation of mitochondrial membrane potential upon helenalin treatment. Moreover, helenalin induces an atypical form of cell death with necrotic features in Bcl-2 overexpressing cells, neither activating classical mediators of apoptosis (caspases, AIF, Omi/HtrA2, Apaf/apoptosome) nor ER-stress mediators (BiP/GRP78 and CHOP/GADD153), nor autophagy pathways (LC3 conversion). In contrast, helenalin was found to inhibit NF-κB activation that was considerably increased in Bcl-2 overexpressing Jurkat cells and promotes cell survival. Moreover, we identified reactive oxygen species (ROS) and free intracellular iron as mediators of helenalin-induced cell death whereas activation of JNK and abrogation of Akt activity did not contribute to helenalin-elicited cell death. Our results highlight the NF-κB inhibitor helenalin as a promising chemotherapeutic agent to overcome Bcl-2-induced cell death resistance.

Pharmacological properties of motion vision in goldfish measured with the optomotor response.

In goldfish, the retinal pathways involved in motion coding have been demonstrated to have an L-cone dominated action spectrum (S. Schaerer, C. Neumeyer, Motion detection in goldfish investigated with the optomotor response is "color blind", Vision Res. 36 (1996) 4025-4034). The neurotransmitters involved in retinal motion coding mechanisms, and the relevance of these retinal motion coding mechanisms for motion perception, are little investigated in fish. In this study, the optomotor response was used to investigate the effect of antagonists on different receptor types for acetylcholine (ACh), GABA, for the dopamine D2-receptor (D2-R) - which is known to modulate the action spectrum in motion coding (C. Mora-Ferrer, K. Behrend, Dopaminergic modulation of photopic temporal transfer properties in goldfish retina investigated with the ERG, Vision Res. 44 (2004) 2067-2081) - and of an agonist for against the mGluR6-receptor (mGluR6) on goldfish motion vision in the photopic range. Blockade of nicotinic ACh-R, GABAa-R and both GABAa- and GABAc-R eliminated the optomotor response completely. Neither a muscarinic ACH-R antagonist, a D2-R antagonist or a mGluR6-agonist affected goldfish motion vision. The pharmacological profile of the goldfish optomotor response resembles the pharmacological profile of direction-selective ganglion cells (DS-GC) described for vertebrate retinas in electrophysiological experiments, e.g. (S. Weng, W. Sun, S. He, Identification of ON-OFF direction-selective ganglion cells in the mouse retina, J. Physiol. 562 (2005) 915-923). This indicates that cells with direction-selective receptive field properties exist in the goldfish retina. It is proposed that these cells provide the input for the full field motion perception in goldfish.