In recent years, natural products gained popularity with their anti-inflammatory and antioxidant effects mediated by chemical compounds within their composition. Study results offering them as palliative therapy options in cancer or as anticancer agents with high levels of cytotoxicity brought a new approach to combine cancer treatment protocols with these products. From a different perspective, edible types of these products are suggested in daily diets due to their potential cancer preventive effects. Our preliminary work was on blueberry extracts (Vaccinium myrtillus) as a main representative of these natural products, and the contents of the extracts were analyzed with liquid chromatography tandem mass spectrometry (LC MS/ MS) to reveal the composition and distribution of polyphenolic compounds within. The most abundant polyphenols detected in V. myrtillus extracts were quercetin, kaempferol, and a phenolic acid, gentisic acid (GA). The compounds were further evaluated on treated HCT-116 cells for their potential anticancer effects by measuring total antioxidant status, total oxidant status, and 8hydroxydeoxyguanosine levels for evaluation of oxidative stress and through protein array analysis and flow cytometric analysis for evaluation of apoptosis. In analysis of oxidative stress parameters, reduced total oxidant levels and reduced oxidative stress index levels were found in cells treated with the compounds in comparison with untreated cells. In apoptosis-related protein profiles, at least twofold reduction in various apoptotic proteins was observed after quercetin and kaempferol treatment, whereas a different profile was observed for GA. Overall, results of this study showed that quercetin and kaempferol have strong cytotoxic, antioxidant, and apoptotic effects, although GA is mostly effective as an antioxidant polyphenol on HCT-116 cells.
Previously, we demonstrated that biotransformation of propolis by some special strains of Lactobacillus plantarum might decrease the allergenic molecules in propolis.In this study, we aimed to investigate the effect of biotransformation of propolis on its antioxidant effect and its protective effect against potassium bromate-induced cancer in human colon cell line. Propolis samples were treated with different solutions (ethanol, polyethylene glycol, and water), and ultrasonication was applied at 40 Hz (5, 10, and 15 minutes) in order to facilitate solvation of solid samples. Fermentations were performed by L. plantarum strains (ISLG-2, ATCC-8014, and Visbyvac). The phenolic content of propolis was determined with liquid chromatography-mass spectrometry/mass spectrometry (LCMS/MS). The antioxidant activity (antioxidant enzymes, lipid peroxidation) and apoptosis markers (caspase 3,8,9, cytochrome-c, tumour necrosis factor-related apoptosis-inducing ligand-R1 and R2 [TRAIL], and apoptosis protease activating factor-1 [APAF-1] levels) were determined in CCD 841-human colon cell line after induction of oxidative stress by potassium bromate. All propolis samples in different solvents induced apoptosis and 4 biotransformed (by L. plantarum ISL-2 strain and L. plantarum ATCC 8014 strain) propolis samples with low allergenic molecules demonstrated similar inductions of apoptosis in CCD841 cell line.In conclusion, reduction of allergenic molecules in propolis via biotransformation did not change the antioxidant and protective effects of propolis, and it is suggested as a potential therapeutic molecule in prevention of colon cancer caused by oxidative stress for all patients.Significance of the study: This study is the first investigation that shows protective effect of propolis against potassium bromate toxicity by means of decreasing lipid peroxidation and reversing the main molecule levels in intrinsic and extrinsic pathway of apoptosis. Biotransformed propolis samples by L. plantarum ISL-2 and ATCC 8014 strain with low allergen molecule content has also the same effect in potassium bromate toxicity in CCD841 colon cell. Our data contributed that propolis as a natural
Propolis has been used for its health benefits, due to high phenolic content. Recently it has been shown that the extraction methods which yielded phenolic molecules, affected the anti-oxidant and anticancer effect of propolis. In our previous study we showed that biotransformation of propolis via Lactobacillus plantarum might increase antioxidative effect. In this study, we investigated the cytotoxic effect of this propolis sample on A375 melanoma cells. The propolis samples were extracted in water. The phenolic molecules were determined with LC MS/MS. Cytotoxicity was evaluated by means of the WST. Water-extracted propolis samples were incubated with L. plantarum (1.5%) in 37 °C for 24 h. A375 cells were treated by propolis with doses of from 25 to 1000 µg/mL, for periods of 24 h, 48 h and 72 h. Cytotoxicity MTT tests were performed. The significantly high phenolic compounds mainly; Quercetine (514 ng/mL), rutin (623 ng/mL), ellagic acid (331 ng/mL), epicatechin (125 ng/mL) were found in propolis samples IC50 values were 412.5 µg/mL (24 h) and 314 µg/mL (48 h) and 353 µg/mL (72 h). In conclusion, our data showed that the cytotoxic effect of biologically transformed propolis which have high content of rutin, quercetin, ellagic acid, epicatechin. Biotransformation might be a useful strategy to increase bioavailability of phenolic molecules in propolis.
In this study, the fruits and leaves of wild blueberries grown naturally in our country will be evaluated by using different infusion and boiling methods. Blueberry teas; leaves, raw fruit, dried and shredded raw fruit, fruit beans and seedless raw fruit of different infusions were used after boiling them for 1 min, 3 min, 5 min, 7 min, 10 min. Phenolic levels were determined by LC MS/MS technique. The antioxidant and activities of all products in the vitro HCT-116 colon cancer cell line were analyzed by spectrophotometric methods. MDA and TEAC were evaluated for antioxidant activity. Cytotoxicity and viability tests were performed by adding WST-8 (Water Soluble Tetrazolium Salt-8) solution. For apoptosis, TRAIL and Apaf-1 ELISA Kit were used for the activation of caspases of intrinsic and extrinsic pathways.
SUMMARYObjective: It has been shown that hydroxytryrosol (HT) has antitumoral, anti-proliferative and apoptotic effects on various cancer cell line types. In this study, we investigated cytotoxic and apoptotic effects of HTon human ovarian cancer cell lines OVCAR-3 and MDAH-2774 and relationship of its cytotoxic effects with phosphatidylinositol 3-kinase/Akt (PI3K/Akt) and extracellular signal-regulated kinase 1/2 (ERK 1/2) signaling pathways. Method: XTT cell proliferation kit, Cell Death Detection Elisa Plus Kit and Human Apoptosis Array test were used to determine the cytotoxic and apoptotic effects of HT in OVCAR-3 and MDAH-2774 cell lines at 24, 48, 72 and 96 hours (h). Effect of HT on PI3K/Akt and ERK 1/2 signaling pathways were investigated by using specific inhibitors of these pathways. Results: IC50 values of HT were found to be 102.3 μM in MDAH-2774 cells at 72 h and 51.5 μM in OVCAR-3 cells at 96 h. Highest apoptotic effect was seen at 100 μM concentration in OVCAR-3 cells at 96 h and at 50 μM at 72 h in MDAH-2774. HT increased the levels of pro-apoptotic molecules including Bad, Bax, active caspase-3, Htra2/Omi and of proapoptotic death receptors TRAIL R1/DR4, TRAIL R2/DR5, FAS/TNFRSF6 and decreased Survivin levels in MDAH-2774 cells. In OVCAR-3 cells, HT decreased Bcl-2, pro-caspase 3 and CIAP-1, CIAP-2, XIAP, Livin, Survivin levels and increased cytochrome-c level. The cell vitality was increased in both of the cell lines as the result of the treatment with HT and FR180204. Conclusions: These results show that HT has potential cytotoxic and apoptotic effect on OVCAR-3 and MDAH-2774 cells. In both cell lines, the cytotoxic effect of HT could be ocur inhibiting through ERK 1/2 signaling pathway.
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