Tabolicproperties [2]. This assumption has not but been confirmed experimentally. The second objective of this study, thus, was to evaluate the antiproliferative properties of FWGE as well as the DMBQ compound (at a concentration equal to that in FWGE) in nine human cancer cell lines.MethodsCell linesHuman malignant cell lines (Table 1) have been routinely cultured in RPMI 1640 medium at 37 and 5 CO2 supplemented with ten (v/v) heat-inactivated fetal calf serum (FCS), 100 U/ml penicillin, one hundred g/ml streptomycin, 2 mmol/l glutamine, 50 mmol/l mercaptoethanol, and 1 non-essential amino acids in final concentrations (Invitrogen Life Technologies GmbH, Karlsruhe, Germany).Drugs and chemicalsFWGE powder from Biropharma, Hungary, was employed. For every experiment, a fresh stock solution containing 100 mg/ml FWGE was prepared with RPMI 1640 medium and passed by way of a 0.two m filter. FWG includes benzoquinones within a concentration of approx. 400 g/g (0.04 ) crude extract as described previously [2]. The antitumor compound DMBQ (molecular weight: 168 g/mol) at 97 purity (Sigma-Aldrich, Germany) was made use of within the same molar concentration of 24 mol/l as described for FWGE (Avemar) [2]. The DMBQ concentration for ten mg/ml FWGE (=24 mol/l) was calculated as follows: [10 mg/ml (10 g/l) x 0.04 ]/Table 1 IC50 values of FWGE for different human cancer cell lines. IC50 values in mg/ml are shown as mean normal deviation (SD) of 3 independent experiments, each performed in triplicate and calculated as described [21]. Cancer cells had been treated for 24 h with unique concentrations of FWGE (0.1, 1.0, ten, 50 mg/ml) in medium with ten (v/v) fetal calf serum (FCS) and cell viability was determined by crystal violet (CV)Cell line MDA-MB-468 ASPC-1b BxPC-b aTissue Adenocarcinoma with the breast [33] Adenocarcinoma of the pancreas [34] Adenocarcinoma on the pancreas [34] Adenocarcinoma of the breast [33] Adenocarcinoma of your stomach [35] Adenocarcinoma with the colon [36] Adenocarcinoma on the breast [37] Adenocarcinoma on the colon [38] Invasive breast ductal carcinoma [37]Mean 3.eight 4.0 4.four 5.five 7.9 ten.9 13.three 15.eight 19.SD 1.77 0.37 0.66 0.08 two.05 6.36 7.41 7.83 17.MDA-MB-231a 23132/87b HT-29a BT-20c HRT-18b MCF-7aCell sources: aCLS: Cell Lines Services GmbH, Eppelheim, Germany (www.clsgmbh.de); bIOP: Institute of Pathology, University of W zburg, Germany.6-Bromo-8-iodoquinolin-2(1H)-one Chemscene cDOG: Division of Obstetrics and Gynaecology, University of W zburg Hospital, GermanyOtto et al.Formula of Fmoc-Gly-OH BMC Complementary and Option Medicine (2016) 16:Page 3 of168 g/mol.PMID:25027343 For each and every experiment, a fresh stock option of 0.1 mg/ml (595 mol/l) DMBQ was set-up in RPMI 1640 medium at 37 for ten min to enhance solubility. Immediately after passing the stock remedy through a 0.two m filter, no DMBQ sediment was observed. A fresh and sterile stock answer of 2.four mmol/l ascorbic acid in RPMI 1640 medium was ready as described [16].Measurement of FWGE effects on cell growthGlucose consumption and lactic acid productionCells (1.504) have been seeded in 200 l culture medium per well into 96-well flat-bottom tissue plates (Greiner bio-one, Germany). The next day, cells had been incubated for 24 h with fresh culture medium containing the final concentrations of 0.1, 1.0, 10, 50 mg/ml FWGE according to published data [179]. The tissue plates have been cultured at 37 in a humidified atmosphere of 5 CO2 in air and cellular viability was determined by crystal violet (CV) staining as described previously [20]. The absorbance (optical density, OD) was me.