Different effects of two cyclic chalcone analogues on cell cycle of Jurkat T cells.
It has been previously shown that the cyclic chalcone analogues E-2-(4'-methoxybenzylidene)- (2) and E-2-(4'-methylbenzylidene)-1-benzusuberone (3) inhibited proliferation of various murine and human tumor cells. In order to gain new insights into the cytotoxic mechanism of the two compounds detection of apoptosis and necrosis of Jurkat T cells exposed to 2 and 3 were performed by flow cytometry using the Annexin V-FITC and propidium iodide double staining method. Analysis of the DNA histograms at 8, 24 and 48 h exposure times showed that near equitoxic doses of 2 and 3 had different effects on the cell cycle of the exposed cells. The immediate (8h) effect of 2 was a remarkable decrease of cells in the G(0)/G(1) phase and increase in the G(2)/M phase. This effect could also be seen in the histogram of cells at the 24h time point. On the contrary, such an effect of 3 could not be observed. At the 24 and 48 h time points accumulation of sub-G(0) (apoptotic and necrotic) and hyperdiploid cells could be detected after both treatments. Incubation of 2 and 3 with reduced glutathione under cell-free conditions indicated spontaneous conjugation (non-redox) reaction at pH 7.4 and pH 9.0. Analyzing the mechanism of action the total thiol content of the cells exposed to compounds 2 and 3 was determined. Compound 2 showed to reduce the total cellular thiol level both under nutrient-free and nutrient-supplemented conditions. Under the latter conditions an increase of the total thiol level of the cells exposed to 3 for 4h could be observed. The different effect of the two compounds on the cellular thiol status might contribute to the different tumor cytotoxicity of the cyclic chalcone analogues 2 and 3. Investigation of antioxidant capacity of the compounds by monitoring time course of the Fenton-reaction initiated in vitro degradation of 2-deoxyribose indicated that both compounds displayed hydroxyl radical scavenger activity. The experiments provide further details of dual--cytotoxic and cytoprotective (chemopreventive)--effects of the compounds.
Rozmer
Zsuzsanna
Z
Institute of Pharmaceutical Chemistry, University of Pécs, P.O. Box 99, H-7602 Pécs, Hungary.
Berki
Timea
T
Perjési
Pál
P
D017302:Annexins; D000970:Antineoplastic Agents / Q000737:chemistry / Q000633:toxicity*; D000975:Antioxidants / Q000494:pharmacology; D001593:Benzyl Compounds / Q000737:chemistry* / Q000633:toxicity*; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D047188:Chalcones / Q000737:chemistry / Q000633:toxicity*; D004396:Coloring Agents; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D003855:Deoxyribose; D005434:Flow Cytometry; D016166:Free Radical Scavengers / Q000494:pharmacology; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide; D017665:Hydroxyl Radical / Q000378:metabolism; D007501:Iron; D019169:Jurkat Cells; D011419:Propidium; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013729:Terpenes / Q000737:chemistry* / Q000633:toxicity*
D017302:Annexins; D000970:Antineoplastic Agents; D000975:Antioxidants; D001593:Benzyl Compounds; D047188:Chalcones; D004396:Coloring Agents; D004273:DNA, Neoplasm; C515537:E-2-(4'-methoxybenzylidene)-benzusuberone; C515536:E-2-(4'-methylbenzylidene)-1-benzusuberone; C045076:Fenton's reagent; D016166:Free Radical Scavengers; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D013729:Terpenes; D017665:Hydroxyl Radical; D011419:Propidium; D003855:Deoxyribose; D006861:Hydrogen Peroxide; D007501:Iron; D005978:Glutathione