esculetin and Leukemia

Esculetin (6,7-dihydrocoumarin) and the flavonoid quercetin (3,5,7,3',4' pentahydroxyflavone) are compounds that could change the balance of redox homeostasis. NB4 leukemia cells were treated with 25 μM quercetin for 24 h and with esculetin at either 100 or 500 μM for different times. Quercetin increased the levels of pro-inflammatory NFkB p65 in the nucleus correspondingly reducing them in the cytosol. The levels of NFkB p65 decreased in the nucleus at high esculetin concentration treatments for long times (19 h), concomitantly increasing the levels of anti-inflammatory NFkB p50 in the nucleus. This could suggest formation of inhibitory p50 homodimers possibly related with anti-inflammatory response. Lipoxygenase expression was reduced either by esculetin or quercetin. A significant increase of Nrf2 in the nucleus of NB4 cells treated with 100 μM esculetin for 19 h was observed. Quercetin increased the levels of Nrf2 in the cytosol reducing them in the nucleus. Superoxide dismutase expression increased in NB4 cells treated with esculetin in contrast with quercetin. All these data support a relevant differential role for NFkB and Nrf2 in anti-inflammatory and redox response when apoptosis was induced by esculetin or quercetin in human leukemia NB4 cells.

Topics: Antioxidants; Apoptosis; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Nucleus; Cell Survival; Humans; JNK Mitogen-Activated Protein Kinases; Leukemia; Lipoxygenase; NF-E2-Related Factor 2; NF-kappa B; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Quercetin; Reactive Oxygen Species; Superoxide Dismutase; Umbelliferones

The coumarin esculetin shows antioxidant action on some cell types, both by scavenging ROS and by decreasing ROS production. We have previously demonstrated the induction of apoptosis by esculetin on NB4 human leukaemia cells by an ill-defined mechanism related to ROS levels. In this work, we analyze the effect of the simultaneous treatment with esculetin and two oxidants to observe the early events in the mechanism of esculetin-induced apoptosis. Our results show that, from the early time of 15 min, esculetin acts synergistically with H

Topics: Antioxidants; Apoptosis; Cell Proliferation; Cell Survival; Humans; Hydrogen Peroxide; Leukemia; Oxidation-Reduction; Oxidative Stress; Reactive Oxygen Species; tert-Butylhydroperoxide; Umbelliferones

In order to overcome chemotherapy resistance, many laboratories are searching for agents that increase the sensitivity of cancer cells to anticancer drugs. Arsenic trioxide (As(2)O(3)) is widely used in treating human acute polymyelocytic leukemia (APL). However, solid tumors and other leukemia cells such as U937 promonocytic leukemia cells are insensitive to As(2)O(3). Esculetin, a coumarin derivative, has previously induced cell cycle arrest and apoptosis of HL-60 cells as well as enhanced taxol-induced apoptosis in HepG2 cells, thereby displaying anticancer potential. In this study, esculetin inhibited proliferation and mitogen activated protein kinases (MAPKs) activation in human leukemia U937 cells. Since inhibitors of MAPKs have modulated the GSH-redox state and enhanced the sensitivity of leukemia cells to As(2)O(3)-provoked apoptosis, we monitored the effect of combining esculetin and As(2)O(3) (2.5 microM) on the GSH level. Our study showed that esculetin, PD98059 (MEK/ERK inhibitor), and SP600125 (JNK inhibitor) similarly enhanced the As(2)O(3)-induced GSH depletion. We found that the As(2)O(3) (2.5 microM) treatment slightly induced apoptosis and the pretreatment of esculetin enhanced the As(2)O(3)-provoked apoptosis significantly. In addition, esculetin enhanced the effect of As(2)O(3) on caspase activation in U937 cells. We compared the combined esculetin and As(2)O(3) treatment to the As(2)O(3) treated alone. The combined esculetin and As(2)O(3) treatment increased Bid cleavage, Bax conformation change and cytochrome C release. The study also indicated that esculetin enhanced the As(2)O(3)-induced lysosomal leakage and apoptosis. Furthermore, pretreatment with N-acetylcysteine (NAC) reduced these enhanced effects. Based on these studies, esculetin enhances the As(2)O(3)-provoked apoptosis by modulating the MEK/ERK and JNK pathways and reducing intracellular GSH levels. GSH depletion led to higher oxidative stress which activated lysosomal-mitochondrial pathway of apoptosis.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Antioxidants; Apoptosis; Arsenic Trioxide; Arsenicals; Blotting, Western; Cell Survival; Fluorescent Antibody Technique; Humans; Leukemia; Oxides; U937 Cells; Umbelliferones

Cytosolic fraction of RBL-1 cells transformed arachidonic acid to 12-HETE in addition to the well-recognized 5-hydroxyeicosatetraenoic acid (5-HETE) in the presence of Ca2+, Mg2+ or Mn2+. The identity of 12-HETE was confirmed by gas chromatography-mass spectrometry. Syntheses of 12-HETE and 5-HETE were catalyzed by separate lipoxygenases, since the formation of each product showed differential sensitivity to inhibitors and temperature. 12-Lipoxygenase from RBL-1 cells was also found to be distinct from the enzyme from platelets in calcium sensitivity.

Topics: Animals; Arachidonate Lipoxygenases; Arachidonic Acid; Arachidonic Acids; Basophils; Blood Platelets; Calcium; Hot Temperature; Imidazoles; Leukemia; Lipoxygenase; Lipoxygenase Inhibitors; Rats; Umbelliferones