Matrix metalloproteinase processing of PTHrP yields a selective regulator of osteogenesis, PTHrP1-17.

Parathyroid hormone-related protein (PTHrP) is a critical regulator of bone resorption and augments osteolysis in skeletal malignancies. Here we report that the mature PTHrP1-36 hormone is processed by matrix metalloproteinases to yield a stable product, PTHrP1-17. PTHrP1-17 retains the ability to signal through PTH1R to induce calcium flux and ERK phosphorylation but not cyclic AMP production or CREB phosphorylation. Notably, PTHrP1-17 promotes osteoblast migration and mineralization in vitro, and systemic administration of PTHrP1-17 augments ectopic bone formation in vivo. Further, in contrast to PTHrP1-36, PTHrP1-17 does not affect osteoclast formation/function in vitro or in vivo. Finally, immunoprecipitation-mass spectrometry analyses using PTHrP1-17-specific antibodies establish that PTHrP1-17 is indeed generated by cancer cells. Thus, matrix metalloproteinase-directed processing of PTHrP disables the osteolytic functions of the mature hormone to promote osteogenesis, indicating important roles for this circuit in bone remodelling in normal and disease contexts.

The prognostic utility of the transcription factor SRF in docetaxel-resistant prostate cancer: in-vitro discovery and in-vivo validation.

BACKGROUND: Docetaxel based therapy is one of the first line chemotherapeutic agents for the treatment of metastatic castrate-resistant prostate cancer. However, one of the major obstacles in the treatment of these patients is docetaxel-resistance. Defining the mechanisms of resistance so as to inform subsequent treatment options and combinations represents a challenge for clinicians and scientists. Previous work by our group has shown complex changes in pro and anti-apoptotic proteins in the development of resistance to docetaxel. Targeting these changes individually does not significantly impact on the resistant phenotype but understanding the central signalling pathways and transcription factors (TFs) which control these could represent a more appropriate therapeutic targeting approach. METHODS: Using a number of docetaxel-resistant sublines of PC-3 cells, we have undertaken a transcriptomic analysis by expression microarray using the Affymetrix Human Gene 1.0 ST Array and in conjunction with bioinformatic analyses undertook to predict dysregulated TFs in docetaxel resistant prostate cancer. The clinical significance of this prediction was ascertained by performing immunohistochemical (IHC) analysis of an identified TF (SRF) in the metastatic sites from men who died of advanced CRPC. Investigation of the functional role of SRF was examined by manipulating SRF using SiRNA in a docetaxel-resistant PC-3 cell line model. RESULTS: The transcription factors identified include serum response factor (SRF), nuclear factor kappa-B (NFκB), heat shock factor protein 1 (HSF1), testicular receptor 2 & 4 (TR2 &4), vitamin-D and retinoid x receptor (VDR-RXR) and oestrogen-receptor 1 (ESR1), which are predicted to be responsible for the differential gene expression observed in docetaxel-resistance. IHC analysis to quantify nuclear expression of the identified TF SRF correlates with both survival from date of bone metastasis (p = 0.003), survival from androgen independence (p = 0.00002), and overall survival from prostate cancer (p = 0.0044). Functional knockdown of SRF by siRNA demonstrated a reversal of apoptotic resistance to docetaxel treatment in the docetaxel-resistant PC-3 cell line model. CONCLUSIONS: Our results suggest that SRF could aid in treatment stratification of prostate cancer, and may also represent a therapeutic target in the treatment of men afflicted with advanced prostate cancer.

Carotenoid content of commonly consumed herbs and assessment of their bioaccessibility using an in vitro digestion model.

Herbs are a rich source of bioactive phytochemicals such as carotenoids, which are known to exert various positive biological effects. However, there is very limited information in the literature regarding the content and bioavailability of carotenoids from commonly consumed herbs. Therefore, the objectives of the present study were first, to determine the carotenoid content of eight herbs namely basil (Ocimum basilicum), coriander (Coriandrum sativum), dill (Anethum graveolens), mint (Metha L.), parsley (Petroselinum crispum), rosemary (Rosmarinus officinalis), sage (Salvia officinalis), and tarragon (Artemisia dracunculus L.); and second, to assess carotenoid bioaccessibility from these herbs using a simulated human in vitro digestion model. Carotenoid bioaccessibility is defined as the amount of carotenoids transferred to micelles after digestion when compared with the original amount present in the food. The content of individual carotenoids varied significantly among the herbs tested. Carotenoid bioaccessibility varied from 0 to 42.8%. Basil and coriander, and their respective micelles, contained the highest levels of beta-carotene, beta-cryptoxanthin, and lutein + zeaxanthin. Our findings show that herbs are rich sources of carotenoids and that these foods can significantly contribute to the intake of bioaccessible carotenoids.

Bioaccessibility, uptake, and transport of carotenoids from peppers (Capsicum spp.) using the coupled in vitro digestion and human intestinal Caco-2 cell model.

Spanish bell peppers (Capsicum annuum L.) and chili peppers sourced from Kenya and Turkey were analyzed for their carotenoid content, bioaccessibility, and bioavailability. The order of total carotenoid content in peppers and their respective micelles was red > green > yellow. In terms of cellular carotenoid transport as a percentage of original food and micelle content, the order was yellow peppers > green > red; however, the opposite trend was seen for the actual amount of total carotenoids transported by Caco-2 cells. Although lutein was generally the most abundant carotenoid in the micelles (496.3-1565.7 microg 100 g(-1)), cellular uptake and transport of beta-carotene were the highest, 8.3-31.6 and 16.8-42.7%, respectively. Hence, the actual amount of carotenoids present in the original food and respective micelles seems to reflect the amount transported by Caco-2 cells. Therefore, color influenced the carotenoid profile, bioaccessibility, and bioavailability of carotenoids rather than pepper type.

Growth inhibitory effects of casein hydrolysates on human cancer cell lines.

The aim of this study was to investigate the effects of unhydrolysed/intact casein and eight different sodium casein hydrolysates (a-h) on the viability and growth of human cancer cell lines. Both human Jurkat T cells and Caco-2 cells were incubated with increasing concentrations of the test compounds (0.5-10% v/v) for 24 h. Cell viability was assessed using the MTT, lactate dehydrogenase (LDH) release and Trypan Blue assays. Cell growth was monitored using the MTT, Trypan Blue and Bromodeoxyuridine (BrdU) proliferation assays. Casein hydrolysates b, c and f had an inhibitory effect on the viability and growth of both cell lines. The casein hydrolysates did not negatively affect the membrane integrity of both Jurkat and Caco-2 cells. In Jurkat cells hydrolysates a and h had an inhibitory effect on DNA synthesis after 24 h, while in Caco-2 cells DNA synthesis was not affected. In conclusion, we found that the different casein hydrolysates had cell-specific effects which target particular functions within the cell. Overall, casein hydrolysates had no effect on membrane integrity while they had varied effects on mitochondrial activity and DNA synthesis in the different cell lines.

Bioactivity of herb-enriched beef patties.

Interest exists in the manufacture of meat products with added functional ingredients to enhance consumer health. Because experimental evidence suggests that many herbs and spices, particularly those of the Lamiaceae family such as Salvia officinalis L. (sage) and Origanum vulgare L. (oregano), possess a wide range of biological and pharmacological activities, they represent promising functional ingredients for incorporation into meat and meat products. The present study aimed to determine the bioactivity of cooked beef patties that were enriched with or without sage or oregano extracts (1,200 microg/g). Cooked beef patties were subjected to an in vitro digestion procedure, and the resulting micelles isolated from the digested meats were added to human intestinal Caco-2 cells. The antioxidant potential (ferric reducing antioxidant power [FRAP] value) of enriched beef patties was significantly higher than the FRAP value of non-enriched beef patties, both before and after in vitro digestion. Cell viability significantly increased following treatment with certain concentrations of the micelle fractions from digested sage- or oregano-enriched beef patties. Pretreatment with micelles derived from sage- or oregano-enriched beef patties did not significantly protect against cell injury or DNA damage induced by H(2)O(2). However, micelles derived from digested sage-enriched beef patties (10% vol/vol) significantly increased cellular reduced glutathione (GSH) content. In addition, micelles derived from both sage- and oregano-enriched beef patties (10% vol/vol) significantly protected against H(2)O(2)-induced GSH depletion. Thus, it appears that sage and oregano exhibit some bioactivity within a meat system. Our findings suggest that herbal extracts have potential as possible functional ingredients in meat products.

Geographical location has greater impact on carotenoid content and bioaccessibility from tomatoes than variety.

The suggested health benefits of consuming tomatoes and tomato-based products have been attributed, in part, to the carotenoids present in these foods. Therefore, the objectives of the present study were to (i) analyse carotenoid content and bioaccessibility from different tomato (Lycopersicon esculentum L.) types namely cherry, plum, round, and certain tomatoes-on-the-vine; and (ii) determine if geographical location (Ireland vs Spain) influenced the content and bioaccessibility of carotenoids in tomatoes of the same variety. Carotenoid bioaccessibility is defined as the amount of ingested carotenoids that, after digestion, are available for absorption by intestinal cells. Differences were seen in carotenoid content and bioaccessibility between the different tomato types tested. For instance, Irish round high-lycopene tomatoes contained the greatest amounts of lycopene and lutein but lowest levels of beta-carotene compared with the other Irish tomatoes. Furthermore, the content and bioaccessibility of carotenoids that were sourced from Ireland and Spain also varied greatly. Spanish tomatoes were generally superior in the content, bioaccessibility, and micelle content of carotenoids. To conclude, our findings suggest that geographical location, rather than the type of tomato, seems to have a more pronounced effect on carotenoid bioaccessibility from tomatoes.

miR-29b expression is associated with disease-free survival in patients with ovarian serous carcinoma.

Micro-RNAs are a group of small noncoding RNAs approximately 22 nucleotides in length. Recent work has shown differential expression of mature micro-RNAs in human cancers. We characterized the alteration in expression of miR-29b in ovarian serous carcinoma. miR-29b expression was analyzed using quantitative stem-loop reverse transcriptase polymerase chain reaction on a set of 50 formalin-fixed, paraffin-embedded ovarian serous carcinoma samples. Protein expression of p53, estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2, Ki-67, and insulinlike growth factor 1 was quantified in the corresponding tissue microarray. The expression profile of miR-29b was correlated with clinicopathological and patient survival data. We provide definitive evidence that miR-29b is down-regulated in a significant proportion of ovarian serous carcinomas and is associated with specific clinicopathological features, most notably high miR-29b expression being associated with reduced disease-free survival.

Lack of genoprotective effect of phytosterols and conjugated linoleic acids on Caco-2 cells.

Much interest has focused on the cholesterol-lowering effects of phytosterols (plant sterols) but limited data suggests they may also possess anti-carcinogenic activity. Conjugated linoleic acids (CLA), sourced from meat and dairy products of ruminant animals, has also received considerable attention as a potential anti-cancer agent. Therefore, the aims of this project were to (i) examine the effects of phytosterols and CLA on the viability and growth of human intestinal Caco-2 cells and (ii) determine their potential genoprotective (comet assay), COX-2 modulatory (ELISA) and apoptotic (Hoechst staining) activities. Caco-2 cells were supplemented with the phytosterols campesterol, beta-sitosterol, or beta-sitostanol, or a CLA mixture, or individual CLA isomers (c10t12-CLA, t9t11-CLA) for 48 h. The three phytosterols, at the highest levels tested, were found to reduce both the viability and growth of Caco-2 cells while CLA exhibited isomer-specific effects. None of the phytosterols protected against DNA damage. At a concentration of 25 microM, both c10t12-CLA and t9t11-CLA enhanced (P<0.05) oxidant-induced, but not mutagen-induced, DNA damage. Neither the phytosterols nor CLA induced apoptosis or modulated COX-2 production. In conclusion, campesterol, beta-sitosterol, beta-sitostanol, c10t12-CLA, and t9t11-CLA were not toxic to Caco-2 cells, at the lower levels tested, and did not exhibit potential anti-carcinogenic activity.

Prosthodontic rehabilitation of hypophosphatasia using dental implants: a review of the literature and two case reports.

There are reports in the literature of the various dental features of hypophosphatasia, especially where it affects the deciduous dentition. The descriptions include both the manifestations of the disorder and the subsequent patterns of tooth loss. There are fewer descriptions of the effects of hypophosphatasia on the permanent dentition and little information on the subsequent prosthodontic management of these patients, particularly in relation to the use of dental implants. The aim of this paper was to review the literature on the dental effects of hypophosphatasia, present two cases and describe how one of those patients, a young adult, was successfully rehabilitated using dental implants. That latter patient's pattern of tooth loss as well as some histological and scanning electron microscopic findings of root cementum from the other case is also described.

Bioactive properties of wood knot extracts on cultured human cells.

Not all felled wood is converted to timber or pulp, with the remaining material being a rich source of relatively unexplored and unexploited potentially novel bioactive compounds. Therefore the potential bioactive effects of two softwood knot (the part of the branch encased in the tree stem) extracts--namely, Pinus banksiana Lamb. (Jack pine) and Picea sitchensis (Bong.) Carr. (Sitka spruce)--were investigated by (1) determining their effects on the viability and antioxidant status of human Jurkat T cells, (2) investigating potential cytoprotective and genoprotective effects against oxidative stress in cultured cells, and (3) assessing their effects on concanavalin A (ConA)-induced interleukin-2 (IL-2) production. Initially, both Jack pine knot and Sitka spruce knot extracts were shown to possess strong antioxidant activity as determined by the ferric reducing antioxidant power assay. When added to Jurkat cells, Jack pine knot extract was more toxic compared with Sitka spruce knot extract, with concentrations that resulted in 50% cell death of 153.0 microg/mL and 376.1 microg/mL, respectively. Supplementation of Jurkat cells with wood knot extracts did not affect their glutathione content or catalase activity. Pretreatment of Jurkat cells with Sitka spruce or Jack pine knot extracts protected against H(2)O(2)-induced cell injury. However, none of the extracts protected against H(2)O(2)-induced DNA damage. Jack pine knots, at a concentration of 30 microg/mL, significantly suppressed ConA-induced IL-2 production. Although total phenol content did not differ between the two extracts, gas chromatography analysis did show variation in the types of constituents present. Further research is warranted to elucidate the selective bioactive properties of these softwood knot extracts.

Cellular transport of lutein is greater from uncooked rather than cooked spinach irrespective of whether it is fresh, frozen, or canned.

Lutein, a carotenoid found in significant levels in spinach, has attracted a great deal of attention owing to its reported function as a shield against the photooxidative effects of blue light. Therefore, the rationale of this study was to examine the effects of various processing and cooking methods on lutein bioavailability from spinach (Spinacia oleracea) using an in vitro digestion procedure coupled with the use of a human intestinal Caco-2 cell model. Fresh, frozen, and canned spinach were analyzed uncooked and after boiling or microwave cooking. Lutein content from the uncooked and cooked digested food (digestate) and appropriate micelles was determined. Micellarized lutein from the spinach samples was adjusted to 0.1 micromol/L and added to Caco-2 cells. Cellular uptake and secretion (cellular transport) of lutein were determined. Our results showed that digestate obtained from uncooked canned spinach had greater lutein content (P < .05) than uncooked fresh or frozen spinach. Microwave cooking, but not boiling, significantly lowered the lutein content of canned spinach digestate and micelles compared with their uncooked counterparts. Interestingly, there were no differences in the micellarization of lutein between the cooking and processing methods. Cellular transport of lutein was greater from uncooked spinach micelles compared with boiled or microwave-cooked spinach. To conclude, although the lutein content of digesta and micelles may have been modified, its micellarization was not significantly affected by any of the cooking or processing methods tested. In general, cellular transport of lutein was greatest in uncooked spinach irrespective of whether the spinach was fresh, frozen, or canned.

Micellarisation of carotenoids from raw and cooked vegetables.

The efficiency of carotenoid micellarisation from plant foods can be used as an effective tool for the initial screening of carotenoid bioavailability. Therefore, the objectives of the present study were to assess the effects of cooking on the micellarisation of beta-carotene, lycopene, beta-cryptoxanthin and lutein from courgette (zucchini), red pepper and tomato; and, to a minor extent, investigate uptake of lutein by Caco-2 cells from micellar fractions obtained from raw and cooked courgettes. Both raw and cooked vegetables were subjected to an in vitro digestion procedure. beta-Carotene levels were significantly decreased in the digesta from each vegetable after boiling, grilling, microwave-cooking, or steaming, however all of the cooking methods enhanced beta-carotene transfer to micelles. Carotenoid micellarisation ranged from 1.7% to 100% depending on the food, carotenoid, and the cooking method tested. Grilling and microwave-cooking were generally the most detrimental on the transfer of xanthophyll carotenoids, namely beta-cryptoxanthin, to the micelles. Caco-2 cells absorbed greater amounts of lutein from the micelles of microwave-cooked courgettes than those that were raw, boiled, grilled, or steamed. Depending on the cooking methods used, carotenoid retention as well as micellarisation varied for each carotenoid among the different vegetables and different carotenoids present in each particular food.

Modulation of cytokine production by plant sterols in stimulated human Jurkat T cells.

The plant sterols campesterol, beta-sitosterol and beta-sitostanol were investigated for potential immunomodulatory effects in Jurkat T cells. Treatments involved supplementing cells with or without concanavalin A (ConA) or phorbol-12-myristate-13-acetate plus ionomycin (PMA+IoM) in the presence or absence of increasing concentrations (10-100 microM) of each plant sterol for 24 h. None of the plant sterols significantly affected mitogen-stimulated IL-4, IL-10 or IFN-gamma production. However, campesterol, beta-sitosterol and beta-sitostanol significantly suppressed mitogen-induced IL-2 production in a dose-dependent manner. Both bisindolylmaleimide-I (BIM-I), a specific protein kinase C (PKC) inhibitor, and the immunosuppressant drug known as Tacrolimus (FK506), an IL-2 inhibitor, prevented mitogen-stimulated IL-2 production in Jurkat cells. Treatment with PMA+IoM alone significantly increased PKC activity and the presence of BIM-I prevented PKC activation by PMA+IoM. Following 24 h treatments, the plant sterols did not affect PMA+IoM-enhanced PKC activity, cellular calcium content or calcineurin activity. Intracellular cyclic 3',5'-adenosine monophosphate (cAMP) levels were significantly reduced by PMA+IoM. The presence of FK506 prevented a PMA+IoM-induced reduction of intracellular cAMP. Likewise the plant sterols behaved in a similar manner as FK506. Our findings suggest that the suppression of IL-2 by the plant sterols was not mediated via PKC inhibition and that their effects occurred possibly via cAMP modulation and/or a calcium/calcineurin-independent pathway.

Effects of plant extracts on antioxidant status and oxidant-induced stress in Caco-2 cells.

Experimental evidence suggests that most herbs and spices possess a wide range of biological and pharmacological activities that may protect tissues against O2-induced damage. The objectives of the present study were: first, to determine the effects of plant extracts on the viability, membrane integrity, antioxidant status and DNA integrity of Caco-2 cells and second, to investigate the cytoprotective and genoprotective effects of these plant extracts against oxidative stress in Caco-2 cells. The plant extracts examined were rosemary (Rosmarinus officinalis L.), oregano (Origanum vulgare L.), sage (Salvia officinalis L.) and echinacea (Echinacea purpurea L.). Cell membrane integrity was assessed by the lactate dehydrogenase release assay. Viability was determined by the neutral red uptake assay (NRUA) and the concentration of compound that resulted in 50 % cell death (IC50) was calculated. Antioxidant status of the cells was assessed by measuring GSH content, catalase activity and superoxide dismutase activity. To examine their cytoprotective and genoprotective effects, Caco-2 cells were pre-treated with each plant extract for 24 h followed by exposure to H2O2. DNA damage was assessed by the comet assay and cell injury was determined by the NRUA. Rosemary was the most toxic (IC50 123 microg/ml) and echinacea the least toxic (IC50 1421 microg/ml). Sage was the only plant extract to affect the antioxidant status of the cells by increasing GSH content. Sage, oregano and rosemary protected against H2O2-induced DNA damage (olive tail moment and percentage tail DNA), whereas protection against H2O2-induced cytotoxicity was afforded by sage only.

Polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is the most common endocrine disorder in women of reproductive age, with levels of between 4 and 10 per cent reported (Dunaif 1995, Norman et al 2002). However, figures for its prevalence vary considerably and estimates of 'true' prevalence must be made with caution since there is no overall consensus on the diagnostic criteria that must be satisfied to make a diagnosis (Ledger and Clark 2003). Giving information and patient education are integral to many nursing roles and as more women are being diagnosed with PCOS, patients are seeking information from nurses as well as other health professionals. This article provides an overview of PCOS, including its clinical features, pathophysiology, long-term health consequences and possible treatments.

Dietary flavonols: chemistry, food content, and metabolism.

The flavonols belong to a large group of compounds called flavonoids, which are diverse in their chemical structure and characteristics. Fruits, vegetables, and beverages such as tea and red wine are major sources of flavonols in the human diet. The daily consumption of flavonols is difficult to estimate because values depend on accurate assessment of feeding habits and flavonol content in foods. Food sources, dietary intakes, and bioavailability of flavonols are strongly influenced by variations in plant type and growth, season, light, degree of ripeness, food preparation, and processing, all of which are discussed. In the past few years, a number of studies on the absorption and metabolism of flavonols in humans have been published and the findings from these studies are reviewed. We do not discuss the health effects of flavonols.

Mechanism of protection by the flavonoids, quercetin and rutin, against tert-butylhydroperoxide- and menadione-induced DNA single strand breaks in Caco-2 cells.

Protection by the flavonoids, quercetin and rutin, against tert-butylhydroperoxide (tert-BOOH)- and menadione-induced DNA single strand breaks was investigated in Caco-2 cells. Both tert-BOOH and menadione induced DNA single strand breaks in a concentration-dependent manner. Pre-incubation of Caco-2 cells with either quercetin or rutin for 24 h significantly decreased the formation of DNA single strand breaks evoked by tert-BOOH (P <.05). Iron chelators, 1,10-phenanthroline (o-Phen) and deferoxamine mesylate (DFO), also protected against tert-BOOH-induced DNA damage, whereas butylated hydroxytoluene (BHT) had no effect. Quercetin, and not rutin, decreased the extent of menadione-induced DNA single strand breaks. DFO and BHT, and not o-Phen, protected against menadione-induced DNA strand break formation (P <.05). From the results of this study, iron ions were involved in tert-BOOH-induced DNA single strand break formation in Caco-2 cells, whereas DNA damage evoked by menadione was far more complex. We demonstrated that the flavonoids, quercetin and rutin, protected against tert-BOOH-induced DNA strand breaks by way of their metal ion chelating mechanism. However, quercetin, and not rutin, protected against menadione-induced DNA single strand breaks by acting as both a metal chelator and radical scavenger.

Cytotoxicity, genotoxicity and oxidative reactions in cell-culture models: modulatory effects of phytochemicals.

Much research effort has focused on the identification of phytochemicals in fruit and vegetables which exert beneficial effects. Our research examines modulatory effects of phytochemicals on cytotoxicity, genotoxicity and oxidative reactions in cell systems. Two examples of our studies are discussed. First, the potential beneficial effects of flavonoids are demonstrated. Flavonoids are reported to exhibit a wide variety of biological effects, including antioxidant and free-radical-scavenging activities. The aim of the study was to determine if flavonoids could protect against H2O2-induced DNA damage, as measured by the comet assay, in Caco-2 and HepG2 cells. Both cell lines were supplemented with increasing concentrations of myricetin, quercetin and rutin for 24 h followed by exposure to H2O2 (50 microM) for 30 min. Exposure to H2O2 for 30 min at 37 degrees C resulted in significant DNA damage and pre-incubation with the flavonoids before H2O2 exposure significantly (P <0.05) protected Caco-2 and HepG2 cells against H2O2-induced DNA damage. Secondly, we illustrate the use of cellular models to study oxysterol-induced toxicity. Oxysterols are generated during the cooking and processing of foods and may be produced endogenously by the oxidation of membrane lipids. Recent findings suggest that oxysterols may modulate cytotoxicity by exerting effects on the induction of apoptosis. 7beta-Hydroxycholesterol (7beta-OHC) and 25-hydroxycholesterol, both of which are commonly found in foods, were investigated for their abilities to induce apoptosis in a human monocytic blood cell line, U937, and in the human hepatoma cell line, HepG2 cells. U937 and HepG2 cells were incubated for up to 48 h with 30 microM oxysterol. 7beta-OHC induced apoptosis in U937 cells as measured by non-random DNA fragmentation, condensed and fragmented nuclei, and the generation of hypodiploid cells. In contrast, oxysterols may induce cell death by a different mechanism in the hepatoma cells, possibly by necrosis.

Lack of effect of the flavonoids, myricetin, quercetin, and rutin, on repair of H2O2-induced DNA single-strand breaks in Caco-2, Hep G2, and V79 cells.

In the present study the effects of three flavonoids on the repair of H2O2-induced DNA strand breaks were investigated in Caco-2, Hep G2, and V79 cells. At the concentrations used, myricetin, quercetin, rutin, and H2O2 did not significantly affect cell viability in all the cell lines. Catalase activity was measured in V79 cells and was found to be considerably lower than activities previously measured in Caco-2 and Hep G2 cells. Cells were exposed to 50 microM H2O2 for 0.5 hour at 37 degrees C. After treatment, DNA strand break repair in H2O2-treated cells was monitored at various time points over a 48-hour period using the alkaline single-cell gel electrophoresis assay. Caco-2 cells repaired faster than Hep G2 cells, which repaired considerably faster than V79 cells. Preincubation with 50 microM quercetin for 24 hours significantly decreased the extent of H2O2-induced DNA single-strand breaks throughout repair time points in Caco-2 cells (p < 0.05), but not in Hep G2 cells. Myricetin (50 microM) and rutin (50 microM) had no effect on repair in Caco-2 and Hep G2 cells. Preincubation for 10 hours with quercetin and rutin, but not myricetin, significantly decreased the initial extent of DNA damage induced by H2O2 in V79 cells (p < 0.05). However, from the results of this study, none of the three flavonoids increased the rate of repair of strand breaks in any of the cell types.