Background Tumors are diseases characterized by uncontrolled cell growth and, in

Background Tumors are diseases characterized by uncontrolled cell growth and, in spite of the progress of medicine over the years, continue to represent a major danger to the health, requiring new treatments. that with its core 4-hydroxy-1,4-benzoquinone, exhibits a potent and selective cytotoxic activity on different tumor cell lines. A series of derivatives 3-hydroxy-1,4-benzochinoni were therefore developed through HU-331 chemical modifications. The purpose of the work is to test the ability of the compounds to induce proliferative inhibition and study the mechanisms of cell death. Methods The antitumor activities were evaluated by analyzing their cytotoxic effects against different human being tumor cell lines. All cell lines tested were plated in 96-multiwell and treated with HU-100-V at different concentrations and cell viability was evaluated byMTT assay. Subsequently via circulation cytometry (FACS) it was possible to assess apoptosis by the system of double labeling with PI and Annexin-V, and the effect of the compounds on ROS formation by measuring the dichlorofluorescein fluorescence. Results The substitution by n-hexyl chain substantially enhanced the bioactivity of the compounds. In details, 2-hexyl-5-hydroxycyclohexa-2,5-diene-1,4-dione (V), 2,5-Dimethoxy-3-hexyl-2,5-cyclohexadiene-1,4-dione (XII) and 2-hydroxy-5-methoxy-3-hexyl-cyclohexa-2,5-diene-1,4-dione (XIII) showed most prominent cytotoxicity against almost human being tumour cell lines. Compound V was further subjected to downstream apoptotic analysis, demostrating a time-dependent pro-apoptotic activity on human melanoma M14 cell collection mediated by caspases activation and poly-(ADP-ribose)-polymerase (PARP) protein cleavage. Conclusions These findings show that 2-hexyl-5-idrossicicloesa-2,5-diene-1,4-dione can be a encouraging compound for the design of a new class of antineoplastic derivatives. Carmen Petronzi, Michela Festa, Antonella Peduto and Maria Castellano: equally contributed equally to this work. present in Suriname forests, exhibited activity towards mutant yeast strains, indicative of their cytotoxicity and potential antitumor activity [2]. Furthermore Kaul and co-workers isolated a known cytotoxic quinine Irisoquin which exhibited cytotoxic properties [3]. In previous reports Muhammad et al. [4] evaluated cytotoxic and antioxidant activities of alkylated benzoquinones from your leaves of instrumentMicroanalyses were carried out on Carlo Erba 1106 elemental analyzer. Biological studies Cell cultureOur experimental models consist of several cell lines derived from human cancers of different histogenesis. The cells were produced in RPMI or DMEM supplemented with warmth inactivated 10% FBS, 20 mM HEPES, 100 U/ml penicillin, 100 g/ml streptomycin, 1% L-glutamine in a humidified atmosphere of 95% air flow/5% CO2 at 37C [16]. Analysis of cell proliferation was performed in the presence of all derivatives on all cell lines seeded in 96-well plates at the different densities depending on the cell type. Pancreas malignancy cell lines ( BXPC3, PANC-1) were plated to the average density of 3,600 cells/ well. Prostate malignancy cell lines (DU145, PC3, LNCAP) were plated to the average VX-689 density of 2,000 cells/ well. Melanoma cell lines (COLO38, A375, M14) were plated to the average density of 1 1,800 cells/ well. Renal malignancy cell lines (A498, VX-689 RXF393, SN12C, 769P) and glioblastoma cell lines ( LN229, U87 MG, U373 MG) were plated to the average VX-689 density of 1 1,900 cells/ well. Breast malignancy cell lines (CG5, MCF-7, MDA-MB 231, MDA-MB 468, MDA-MB 436 ) were plated to the average density of 3,100 cells/ well. VX-689 After 24 h incubation at 37C, the cells were Rabbit polyclonal to DUSP13 treated with increasing concentrations of compounds (0,037-50 M). Cells were incubated under these conditions for 72 h. MTT bioassay Human malignancy cells (3 103) were plated in 96-well culture plates in 90 L of culture medium and incubated at 37C in humidified atmosphere of 5% CO2. The day after, 10 L aliquot of serial dilutions of compounds (1C50 M) was added to the cells and incubated for 72 h. The cell viability was assessed with MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] method [17]. After 72 h of treatment with derivatives MTT answer 5 mg/ml in PBS was added to each well. The plates were then incubated at 37C for an additional 4 h to allow MTT to form formazan crystals by reacting with metabolically active cells. The formazan crystals were solubilized in a 1N isopropanol/HCl 10% answer at 37C, on a shaking table for 20 min. The absorbance values of the solution in each well were measured at 570 nm using a micro plate reader. Cell viability was determined by the formula:as previously reported [18]. All MTT experiments were performed in quadruplicated and repeated at least three occasions. Data are as mean standard deviation (SD). Each IC50 imply value was obtained from four impartial experiments. 15 M). The structure-activity relationship studies regarding the first series of compounds revealed that the n-hexyl chain in position 5 of the hydroxy-quinone ring was fundamental for the anticancer activity (compounds II, IV and V), in fact compounds I and III, which lacked of the alkyl chain, were completely inactive. At the same time, the switch of position of alkyl chain was VX-689 clearly detrimental (VI, VII and VIII V). No relevant influence on.

Comments are closed.