Genistein Enhances TRAIL-Mediated Apoptosis Through the Inhibition of XIAP and DcR1 in Colon Carcinoma Cells Treated with 5-Fluorouracil.

Objectives: Colorectal cancer is one of the most common cancers worldwide. However, surgical intervention and chemotherapy provide only limited benefits for the recovery and survival of patients. The anticarcinogenic effect of genistein has attracted attention because epidemiological studies have shown that soybean consumption is associated with a decrease in the incidence of cancer. There are limited studies on the effects of genistein in colorectal carcinoma cells. We aimed to investigate the cytotoxic, genotoxic, and apoptotic effects of genistein in SW480 and SW620 colon adenocarcinoma cells treated with 5-fluorouracil, the basis of chemotherapy, and the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) ligand, the mediator of apoptosis, both alone and in combination. Materials and Methods: Cytotoxicity and genotoxicity were determined by MTT and comet assays, respectively. The apoptotic effects were evaluated by reverse transcription-polymerase chain reaction assay, with the additional use of Annexin V FITC, mitochondrial membrane potential (MMP), caspase 3, 8, and 9 activity, and reactive oxygen species (ROS) assay kits. Results: According to our findings, genistein, 5-fluorouracil, and TRAIL had synergistic apoptotic effects because of DR5 upregulation, ROS production, and DNA damage, which were mediated by increased caspase-8, and -9 activity and decreased MMP. Conclusion: The applied combinations of these compounds may contribute to the resistance problem that may occur in treating colorectal cancer, with a decrease in DcR1 and XIAP genes.

Assessment of the cytotoxic, genotoxic, and apoptotic potential of flurbiprofen in HeLa and HepG2 cell lines.

In the literature, the anticancer potential of flurbiprofen isn't fully understood. In this study, the cytotoxic, genotoxic, and apoptotic effects of flurbiprofen were evaluated in human cervical and liver cancer cells. Cytotoxicity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and it was observed that cytotoxicity increased in a concentration- and time-dependent manner. Genotoxicity was determined using alkaline Comet assay. DNA damage increased in a concentration-dependent manner. Early apoptosis was evaluated using real-time polymerase chain reaction, and it was found that apoptotic gene levels increased while antiapoptotic gene levels decreased. Late apoptosis and cell cycle analyzes were determined using flow cytometry. No evidence of late apoptosis was observed, and no significant arrest was found in the cell cycle. In conclusion, it seems that flurbiprofen has a cytotoxic, genotoxic, and apoptotic effects in both human cancer cell lines. Moreover, the findings indicate that flurbiprofen is effective at the gene level and induces apoptosis with an intracellular pathway.

Evaluation of the Methylation and Acetylation Profiles of Dinitroaniline Herbicides and Resveratrol on the V79 Cell Line.

OBJECTIVES: Herbicides are among the most widely used pesticide compounds for plant growth control worldwide. Risk assessment of the dinitroaniline-derived herbicides pendimethalin and trifluralin is important for foodborne or other means of exposure. In this study, we aimed to evaluate the methylation and acetylation profiles of pendimethalin and trifluralin, which we have high levels of exposure to in various ways. Furthermore, we also determined the protective effect of resveratrol, an antioxidant compound, against the possible toxic effects of these pesticides. MATERIALS AND METHODS: The effects of pendimethalin and trifluralin alone (25, 50, 100 µM) and in combination with resveratrol (100 µM) on DNA methyltransferase (DNMT1) 1, 3a, and 3b; and histone deacetylase (HDAC) 1 and HDAC3 gene expression were evaluated by real-time polymerase chain reaction. RESULTS: According to the results, pendimethalin caused a significant decrease in DNMT1, 3a, 3b and HDAC expressions at all concentrations, whereas HDAC1 and 3 expression was increased at the concentration of 25 µM, when applied together with resveratrol. There were no changes in DNMT1 or 3b expression levels. Unlike pendimethalin, trifluralin increased DNMT1 expression in a concentration-dependent manner. While DNMT3a and DNMT3b expression levels increased significantly, HDAC1 and 3 expression levels did not change significantly. The expression levels of HDAC1 and HDAC3 increased at all concentrations of trifluralin combination with resveratrol. Moreover, DNMT levels increased at the concentrations of 50 and 100 µM. CONCLUSION: Epigenetic gene expression results showed that pendimethalin and trifluralin might cause tissue function loss and chromosome damage as a result of direct effects on cell viability by causing expression level changes in all studied genes. It can also be concluded that the changes that occur in gene expression may induce tumor development. Further studies are needed to elucidate the possible toxicity mechanisms of these herbicides, considering the relationship between epigenetic changes and various diseases.

Preventive role of Pycnogenol® against the hyperglycemia-induced oxidative stress and DNA damage in diabetic rats.

Diabetes mellitus, a complex progressive metabolic disorder, leads to some oxidative stress related complications. Pycnogenol® (PYC), a plant extract obtained from Pinus pinaster, has been suggested to be effective in many diseases including diabetes, cancer, inflammatory and immune system disorders. The mechanisms underlying the effects of PYC in diabetes need to be elucidated. The aim of this study was to determine the effects of PYC treatment (50 mg/kg/day, orally, for 28 days) on the DNA damage and biochemical changes in the blood, liver, and kidney tissues of experimental diabetic rats. Changes in the activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase enzymes, and the levels of 8-hydroxy-2'-deoxyguanosine, total glutathione, malondialdehyde, insulin, total bilirubin, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transferase, high density lipoprotein, low density lipoprotein, total cholesterol, and triglyceride were evaluated. DNA damage was also determined in the whole blood cells and the liver and renal tissue cells using the alkaline comet assay. PYC treatment significantly ameliorated the oxidative stress, lipid profile, and liver function parameters as well as DNA damage in the hyperglycemic rats. The results show that PYC treatment might improve the hyperglycemia-induced biochemical and physiological changes in diabetes.

Effects of cinnamic acid on complications of diabetes

Background/aim: Diabetes mellitus (DM) is a major health problem worldwide. Cinnamic acid (CA) and its derivatives are synthesized in plants and increasing attention has been given to them in recent years due to the high number of beneficial health properties attributed to their consumption. The aim of this study was to investigate the effects of CA on streptozotocin-induced diabetes in Wistar albino rats. Materials and methods: DNA damage was evaluated in the blood, liver, and kidney cells of rats by the alkaline comet assay. Oxidative stress parameters such as catalase, superoxide dismutase, glutathione reductase, glutathione-S-transferase, and glutathione peroxidase activities and 8-hydroxy-2-deoxyguanosine, total glutathione, and malondialdehyde levels; biochemical parameters including insulin, total bilirubin, and BCA protein levels; hepatic enzyme levels such as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase; and lipid profile parameters including high-density lipoprotein, low-density lipoprotein, total cholesterol, and triglyceride levels were also evaluated. Results: DM caused genotoxic damage and alterations in lipid profiles, oxidative stress parameters, and hepatic enzymes levels. CA treatment ameliorated these effects. Conclusion: It seems that CA might have a role in the prevention of the complications of diabetes.

In vitro genotoxicity assessment of dinitroaniline herbicides pendimethalin and trifluralin.

Pendimethalin and trifluralin are widely used dinitroaniline herbicides. Both compounds can be found as residue levels in agricultural products. This study was conducted in order to provide necessary information for the risk assessment of pendimethalin and trifluralin. In this study, reactive oxygen species (ROS) levels were measured to examine the potential of both compounds to induce oxidative damage in Chinese hamster lung fibroblast (V79) cells. Also, the genotoxic effects of pendimethalin and trifluralin at the concentration range of 1-500 µM was determined. Single cell gel electrophoresis (comet) and micronucleus assays were used on human peripheral lymphocytes and V79 cells for the genotoxicity assessment. The cell viability of two dinitroaniline herbicides were determined by the use of neutral red uptake assay on V79 cells. IC50 values were determined as 66 µM and 128 µM for pendimethalin and trifluralin, respectively. They significantly increased ROS levels on V79 cells for 1-24 h. Both herbicides significantly induced the DNA damage and showed genotoxicity on lymphocytes and V79 cells. Micronucleus frequency increased significantly after pendimethalin and trifluralin treatment of the lymphocytes and V79 cells. Therefore, we concluded that both of the herbicides induced the genotoxicity through the activation of oxidative stress pathway and chromosomal damage.

Protective Effects of Ursolic Acid in the Kidneys of Diabetic Rats.

OBJECTIVES: Diabetes, a heteregenous metabolic and chronic disease, is a growing health problem in most countries. It has been claimed that diabetes is associated with the increased formation of free radicals and decreased in antioxidant potential. Oxidative stress formed in diabetes may cause DNA damage in the tissues. Ursolic acid, a well-known pentacylic triterpene, is commonly used in traditional Chinese medicine due to its beneficial health effects such as antioxidant, anticancer, and antiulcer properties. The aim of this study was to investigate the effects of ursolic acid in the kidneys of Wistar albino rats with streptozotocin-induced diabetes. MATERIALS AND METHODS: DNA damage was evaluated in the kidney cells of rats using alkaline comet assays. Oxidative stress parameters such as CAT, SOD, GR, and GSH-Px enzyme activities and total GSH and MDA levels were also evaluated. RESULTS: Ursolic acid treatment was found to significantly decrease DNA damage, GR enzyme activities, and MDA levels, and significantly increase GSH levels and CAT, SOD and GSH-Px enzyme activities in diabetic rats. CONCLUSION: According to our results, it seems that ursolic acid may be beneficial against diabetes-induced renal damage.

The Apoptotic and Anti-apoptotic Effects of Pendimethalin and Trifluralin on A549 Cells In Vitro.

OBJECTIVES: Pendimethalin and trifluralin are commonly used in many countries to control broadleaf weeds and grassy weed species because of their inhibitor effects on growth and cell division. In this study, we examined the apoptotic and anti-apoptotic potentials of pendimethalin and trifluralin on A549 human non-small lung cancer cells with several concentrations in vitro. MATERIALS AND METHODS: The expression levels of apoptosis-related genes BCL-2, BAX, CAS3, CAS9, P53, BIRC, and PPIA were examined using quantitative RT-PCR after 24 h treatment of 1, 10, 50, 100 and 500 µM pendimethalin and trifluralin. RESULTS: The effects of pendimethalin were found more repressive than trifluralin on all studied concentrations. Twenty-four hours' exposure with 100 µM pendimethalin and trifluralin altered the gene expressions, suppressing apoptosis and allowing cancer cells to grow and proliferate. CONCLUSION: Care should be taken not to exceed the permissible values and residue limits in food during pendimethalin and trifluralin use in order to reduce the possible carcinogenic effects on humans.

d-limonene ameliorates diabetes and its complications in streptozotocin-induced diabetic rats.

It is known that diabetes causes some complications including alterations in lipid profile, hepatic enzyme levels but also it causes oxidative stress. Limonene, a major component of Citrus oils, has important health beneficial effects in lowering the level of oxidative stress due to its antioxidant activity. The aim of this study was to investigate the effects of D-limonene on streptozotocin (STZ)-induced diabetes in Wistar albino rats. For this purpose, DNA damage was evaluated by alkaline comet assay. Changes in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and glutathione peroxidase (GSHPx) and the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), total glutathione (GSH), malondialdehyde (MDA), insulin, total bilirubin and BCA protein, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT), high density lipoprotein (HDL), low density lipoprotein (LDL), total cholesterol and triglyceride were also evaluated. D-limonene treatment was found to significantly decrease DNA damage, GR enzyme activities and MDA levels and significantly increase GSH levels and CAT, SOD and GSH-Px enzyme activities and altered lipid and liver enzyme parameters in diabetic rats. According to our results, it seems that D-limonene might have a role in the prevention of the complication of diabetes in rats.

Assessment of Genotoxic Effects of Pendimethalin in Chinese Hamster Over Cells by the Single Cell Gel Electrophoresis (Comet) Assay.

OBJECTIVES: Pendimethalin (N-(1-ethylpropyl)-3,4-dimethyl-2,6-dinitrobenzeneamine) is a dinitroaniline herbicide compound which selectively controls weeds. It is a cell division and growth inhibitor. It descends plants in a short time after seedling. It is a soil and water pollutant due to the widespread use of formulations in Turkey and around the world. Pendimethalin is manufactured in and imported by Turkey. Pendimethalin is a slightly toxic compound that is classified in toxicity class 3 by the United States Environmental Protection Agency (USEPA). Even though it is classified as group C (human possible carcinogen) compound by the USEPA, there are limited number of studies about its genotoxic effects. The aim of this study was to evaluate in vitro genotoxic effects of different concentrations of pendimethalin in Chinese hamster over (CHO) cells by the single cell gel electrophoresis (comet) assay. MATERIALS AND METHODS: The cells are incubated with 1, 10, 100, 1000 and 10000 µM concentrations of pendimethalin for 30 min at 37°C and DNA damage was compared with CHO cells untreated with pendimethalin. 50 µM hydrogen peroxide was used as positive control. RESULTS: No significant cytotoxic effects were observed within the concentration ranges studied. The DNA damage in CHO cells was significantly increased in the pendimethalin concentrations of 1, 100, 1000 and 10000 µM, however, a significant decrease was observed in 10 µM pendimethalin concentration. CONCLUSION: Our results show that 1-10000 µM concentrations of pendimethalin induce DNA damage in CHO cells, which was assessed by comet assay.