ascorbic-acid and Cell-Transformation--Neoplastic

Cancer remains one of the most feared and dreaded diseases in this era of modern medicine, claiming the lives of many, and affecting the quality of life of several others around the globe despite major advances in the diagnosis, treatment, palliative care and the immense resources invested into cancer research. While research in cancer has largely focused on the neoplasm/tumor and the cancerous cells that make up the tumor, more recently, the existence, proliferation, differentiation, migration and invasion of cancer stem cells (CSCs) and the role that CSCs play in tumor initiation, progression, metastasis, drug resistance and relapse/recurrence of the disease has gained widespread interest in cancer research. Although the conventional therapeutic approaches such as surgery, chemotherapy and radiation therapy are effective cancer treatments, very often these treatment modalities fail to target the CSCs, which then later become the source of disease recurrence. A majority of the anti-cancer agents target rapidly dividing cancer cells and normal cells and hence, have side effects that are not expected. Targeting CSCs remains a challenge due to their deviant nature with a low proliferation rate and increased drug resistance mechanism. Ascorbic acid/Vitamin C (Vit.C), a potent antioxidant, is a cofactor for several biosynthetic and gene regulatory enzymes and a vital contributor to immune defense of the body, and was found to be deficient in patients with advanced stages of cancer. Vit.C has gained importance in the treatment of cancer due to its ability to modulate the redox status of the cell and influence epigenetic modifications and significant roles in HIF1α signaling. Studies have reported that intravenous administration of Vit.C at pharmacological doses selectively kills tumor cells and targets CSCs when administered along with chemotherapeutic drugs. In the current article, we provide an in-depth review of how Vit.C plays an important role in targeting CSCs and its possible use as an adjuvant, neoadjuvant or co-treatment in the treatment of cancers.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Ascorbic Acid; Cell Differentiation; Cell Transformation, Neoplastic; Combined Modality Therapy; Drug Resistance, Neoplasm; Humans; Neoplasms; Neoplastic Stem Cells; Signal Transduction

RAS protein family members (KRAS4A, KRAS4B, HRAS and NRAS) function as GDP-GTP-regulated on-off switches, which regulate cytoplasmic-nuclear signaling networks ruling diverse normal cellular processes. Constitutive activating mutations in RAS genes are found in up to 30% of human cancers, and remarkably, the oncogenic Ras mutations and mutations in other components of Ras/MAPK signaling pathways seem to be mutually exclusive in most tumors, pointing out that deregulation of Ras-dependent signaling is an essential requirement for tumorigenesis. Up to 30% of solid tumors are known to have a mutated (abnormal) KRAS gene. Unfortunately, patients harboring mutated KRAS CRC are unlikely to benefit from anti-EGFR therapy. Moreover, it remains unclear that patients with KRAS wild-type CRC will definitely respond to such therapies. Although some clinically designed-strategies to modulate KRAS aberrant activation have been designed, all attempts to target KRAS have failed in the clinical assays and K-RAS has been assumed to be invulnerable to chemotherapeutic attack. Recently, different encouraging publications reported that ascorbate may have a selective antitumoral effect on KRAS mutant cancer cells. In this review we aim to describe the prevalence and importance of KRAS mutation in cancer and associated problems for the clinical handling of patients harboring these tumors. We highlight the role of mutated KRAS in boosting and keeping the tumor associated aberrant cell metabolism stating that further in-depth studies on the molecular mechanism of ascorbate to bypass mutated KRAS-related metabolic alterations may constitute a new pathway to design novel molecules in order handle tumor resistance to anti EGFR-therapies.

Topics: Animals; Antineoplastic Agents; Ascorbic Acid; Biomarkers, Tumor; Cell Transformation, Neoplastic; Drug Discovery; Energy Metabolism; Gene Expression Regulation, Neoplastic; Humans; Molecular Targeted Therapy; Mutation; Neoplasms; ras Proteins; Signal Transduction

The creatine/creatine kinase (CK) system plays a key role in cellular energy buffering and transport. In vertebrates, CK has four isoforms expressed in a tissue-specific manner. In the process of creatine biosynthesis several other important metabolites are formed. The anticancer effect of creatine had been reported in the past, and recent literature has reported low creatine content in several types of malignant cells. Furthermore, creatine can protect cardiac mitochondria from the deleterious effects of some anticancer compounds. Previous work from our laboratory showed progressive decrease of phosphocreatine, creatine and CK upon transformation of skeletal muscle into sarcoma. It was convincingly demonstrated that prominent expression of creatine-synthesizing enzymes L-arginine: glycine amidinotransferase and N-guanidinoacetate methyltransferase occurs in sarcoma, Ehrlich ascites carcinoma and sarcoma 180 cells; whereas, both these enzymes are virtually undetectable in skeletal muscle. Creatine transporter also remained unaltered in malignant cells. The anticancer effect of methylglyoxal had been known for a long time. The present work shows that this anticancer effect of methylglyoxal is significantly augmented in presence of creatine. On creatine supplementation the effect of methylglyoxal plus ascorbic acid was further augmented and there was no visible sign of tumor. Moreover, creatine and CK, which were very low in sarcoma tissue, were significantly elevated with the concomitant regression of tumor.

Topics: Amidinotransferases; Animals; Antineoplastic Agents; Ascorbic Acid; Cell Transformation, Neoplastic; Creatine; Creatine Kinase; Guanidinoacetate N-Methyltransferase; Humans; Membrane Transport Proteins; Mice; Muscle Neoplasms; Muscle, Skeletal; Pyruvaldehyde; Sarcoma

Although ascorbic acid is an important water-soluble antioxidant and enzyme cofactor in plants and animals, humans and some other species do not synthesize ascorbate due to the lack of the enzyme catalyzing the final step of the biosynthetic pathway, and for them it has become a vitamin. This review focuses on the role of ascorbate in various hydroxylation reactions and in the redox homeostasis of subcellular compartments including mitochondria and endoplasmic reticulum. Recently discovered functions of ascorbate in nucleic acid and histone dealkylation and proteoglycan deglycanation are also summarized. These new findings might delineate a role for ascorbate in the modulation of both pro- and anti-carcinogenic mechanisms. Recent advances and perspectives in therapeutic applications are also reviewed. On the basis of new and earlier observations, the advantages of the lost ability to synthesize ascorbate are pondered. The increasing knowledge of the functions of ascorbate and of its molecular sites of action can mechanistically substantiate a place for ascorbate in the treatment of various diseases.

Topics: Animals; Anticarcinogenic Agents; Antioxidants; Ascorbic Acid; Cell Transformation, Neoplastic; Dealkylation; Endoplasmic Reticulum; Glypicans; Histones; Humans; Hydroxylation; Mitochondria; Nucleic Acids; Organelles; Oxidation-Reduction; Proteoglycans; Scurvy; Vitamins

Free radicals can affect the genetic material of cells, causing its gradual impairment and mutation. An accumulation of mutations in certain genes might lead to neoplasmic transformations of the cells and to cancer development. The deteriorative effects of free radicals are counteracted by the antioxidant vitamins A, C, and E that quench free radical reactions. Fruits and vegetables are excellent sources of antioxidant vitamins. The following article attempts a short review of the current knowledge about the influence of vitamins A, C, and E on oxidative damage to DNA, the activity of some transcription factors, and the expressions of certain genes. The aim of this review is to answer the question whether a diet rich in vitamins can protect against cancer.

Topics: Animals; Antioxidants; Ascorbic Acid; Carotenoids; Cell Transformation, Neoplastic; DNA Damage; DNA Repair; Fruit; Humans; Mice; Neoplasms; Oxidation-Reduction; Rabbits; Reactive Oxygen Species; Vegetables; Vitamin A; Vitamin E; Vitamins

Nickel compounds are known to cause respiratory cancer in humans and induce tumors in experimental animals. The underlying molecular mechanisms may involve genotoxic effects; however, the data from different research groups are not easy to reconcile. Here, we challenge the common premise that direct genotoxic effects are central to nickel carcinogenesis and probably to that of other metals. Instead, we propose that it is formation of metal complexes with proteins and other molecules that changes cellular homeostasis and provides conditions for selection of cells with transformed phenotype. This is concordant with the major requirement for nickel carcinogenicity, which is prolonged action on the target tissue. If DNA is not the main nickel target, is there another unique molecule that can be attacked with carcinogenic consequences? Our recent observations indicate that ascorbate may be such a molecule. Nickel depletes intracellular ascorbate, which leads to the inhibition of cellular hydroxylases, manifested by the loss of hypoxia-inducible factor (HIF)-1alpha and -2alpha hydroxylation and hypoxia-like stress. Proline hydroxylation is crucial for collagen and extracellular matrix assembly as well as for assembly of other protein molecules that have collagen-like domains, including surfactants and complement. Thus, the depletion of ascorbate by chronic exposure to nickel could be deleterious for lung cells and may lead to lung cancer. Key words: ascorbate, carcinogenesis, collagens, extracellular matrix, hypoxia-inducible transcription factor, metals, nickel, protein hydroxylation.

Topics: Animals; Antioxidants; Ascorbic Acid; Cell Transformation, Neoplastic; DNA Damage; Homeostasis; Humans; Hydroxylation; Lung Neoplasms; Nickel; Proline; Protein Binding; Rats

Topics: Animals; Anticarcinogenic Agents; Antioxidants; Ascorbic Acid; beta Carotene; Carotenoids; Case-Control Studies; Cell Transformation, Neoplastic; Diet; Free Radicals; Guinea Pigs; Humans; Incidence; Lung Neoplasms; Melanoma; Mice; Mice, Hairless; Models, Chemical; Neoplasms; Neoplasms, Radiation-Induced; Oxygen; Partial Pressure; Prospective Studies; Reactive Oxygen Species; Retrospective Studies; Selenium; Singlet Oxygen; Skin Neoplasms; Smoking; Structure-Activity Relationship; Ultraviolet Rays; Vegetables; Vitamin E

Topics: Ascorbic Acid; Cell Division; Cell Transformation, Neoplastic; Gastric Mucosa; Gastritis; Helicobacter Infections; Helicobacter pylori; Humans; Stomach; Stomach Neoplasms

Topics: Animals; Antineoplastic Agents; Apoptosis; Ascorbic Acid; Cell Division; Cell Survival; Cell Transformation, Neoplastic; Cricetinae; Humans; Neoplasm Proteins; Neoplasms; Tumor Cells, Cultured

Topics: Animals; Antioxidants; Ascorbic Acid; beta Carotene; Carcinogens; Carotenoids; Cell Transformation, Neoplastic; DNA, Neoplasm; Drug Synergism; Humans; Neoplasms, Experimental; Tumor Cells, Cultured; Vitamin E

Vitamins A (retinol, retinoids), beta-carotene (provitamin A), E (alpha-tocopherol), and C (ascorbic acid) are used in experimental, clinical and epidemiological studies for cancer chemoprevention and treatment. The cellular and metabolic effects are depending on the dose used, duration of exposure, and cancer cell type. Despite recent advances, the anticarcinogenic mechanisms remain as yet unknown. Studies regarding the role of vitamins A, beta-carotene, E and C in cancer cell biology and metabolism are of critical importance for their use in cancer treatment. Autoradiographic, ultrastructural and cell surface studies demonstrated that vitamins A, E and C are strong regulator factors of cancer cell differentiation, cell regression, membrane biogenesis, DNA, RNA, protein, and collagen synthesis, as well as transformation of precancer cells into cancer cells. These vitamins exert cytotoxic and cytostatic effects, and may reverse the cancer cell to the normal phenotype. Interrelation of vitamins A, E and C with oncogenes and growth factors play an important role in cancer cell biology. The data presented in this review can provide new insights for the understanding of anticarcinogenic mechanisms, and a rationale for the use of vitamins A, E and C in cancer chemo-prevention and treatment.

Topics: Animals; Anticarcinogenic Agents; Ascorbic Acid; beta Carotene; Carotenoids; Cell Transformation, Neoplastic; Humans; Neoplasms; Vitamin A; Vitamin E

Many studies have now indicated that micronutrients, such as the carotenoids, tocopherols, and ascorbic acid, can prevent mutagenesis, genotoxic effects, or malignant transformation in bacteria and mammalian tissue, either in cell culture or in organ culture. In addition, multiple papers report that some of these micronutrients act as anticarcinogenic agents in animals treated with either ultraviolet light, ultraviolet light with chemicals, or chemical carcinogens alone. With respect to the carotenoids, early experiments used pharmacological doses of carotenoids, but more recent reports indicate that relatively small doses can be effective. Inasmuch as these effects are seen with both provitamin A and nonprovitamin A carotenoids, it would appear that these effects are intrinsic to the carotenoid molecule, and not due to the metabolic conversion to retinoids. Partially on the basis of these observations, it has been suggested that the micronutrients may function as chemopreventive agents for reducing the risk of cancer in humans. Numerous human intervention studies are underway to test this hypothesis.

Topics: Animals; Antimutagenic Agents; Ascorbic Acid; Carotenoids; Cell Transformation, Neoplastic; Cells, Cultured; DNA; DNA, Bacterial; Humans; Mutagenesis; Mutagenicity Tests; Vitamin E

Epidemiological studies have shown that diets rich in one or more antioxidant nutrients may reduce the risk of cancers of the lung, uterine cervix, mouth, and gastrointestinal tract. Study of premalignant lesions offers a comparatively expedient approach to identifying and evaluating the efficacy of the cancer chemopreventive components of foods. Some recent findings suggest roles for beta-carotene and/or vitamin C in reversing or reducing the risk of cervical dysplasia and oral leukoplakia. There are some indications that vitamin C and beta-carotene may reduce the risk of atrophic gastritis and gastric cancer. Additional epidemiological and molecular biology studies and clinical intervention trials using premalignant lesions as the marker of specific cancer risks should become an important component of future research in the area of cancer chemoprevention.

Topics: Ascorbic Acid; beta Carotene; Carotenoids; Cell Transformation, Neoplastic; Diet; Female; Humans; Neoplasms; Precancerous Conditions; Vitamin E

Rodent and human cells in culture, transformed in vitro by radiation or chemicals into malignant cells, afford us the opportunity to probe into early and late events in the neoplastic process at a cellular and molecular level. Transformation can be regarded as an abnormal expression of cellular genes. The initiating agents disrupt the integrity of the genetic apparatus altering DNA in ways that result in the activation of cellular transforming genes (oncogenes) during some stage of the neoplastic process. Events associated with initiation and promotion may overlap to some degree, but in order for them to occur, cellular permissive conditions prevail. Permissive and potentiating factors include free radicals, and thyroid hormone, and inadequate antioxidants. Protective factors which suppress the carcinogenic process include enzymatic and dietary antioxidants. These are constitutive under normal circumstances and can be induced under conditions of oxidative stress produced by a wide range of carcinogens.

Topics: Animals; Antioxidants; Ascorbic Acid; Cell Transformation, Neoplastic; Drug Synergism; Free Radicals; Humans; Selenium; Vitamin E

The data on the influence of antioxidant vitamins (E and C) on the growth of induced and transplantable tumours in animals and on the development of cancer process in man as well as on the course of chemo- and radiotherapy are analyzed; a degree of vitamin provision of the tumour-bearing organism is discussed. Special attention is paid to optimal doses of the above mentioned vitamins. The necessity of combined application of antioxidant vitamins in cancer therapy is established.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Ascorbic Acid; Ascorbic Acid Deficiency; Cell Transformation, Neoplastic; Dose-Response Relationship, Drug; Drug Evaluation; Drug Synergism; Humans; Lipid Metabolism; Lipid Peroxides; Mice; Neoplasms; Neoplasms, Experimental; Nitrosamines; Oxidation-Reduction; Oxidative Phosphorylation; Rabbits; Rats; Tissue Distribution; Vitamin E; Vitamin E Deficiency

The effects of vitamins A, C, and E and of selenium on carcinogenesis are briefly summarized and updated. These vitamins and minerals were selected because they have been studied extensively in recent years with a variety of carcinogenesis models. The consumption of vitamin A and its precursors (carotenoids) has been negatively correlated with cancer at a number of sites, particularly the lung. Animal investigations on vitamin A involvement in carcinogenesis have generally been of three types: those assessing the effect of vitamin A deficiency, the effect of excess vitamin A, or the effect of supplementation with synthetic analogs of vitamin A. Vitamin A deficiency had no effect on salivary gland carcinogenesis, enhanced urinary bladder, lung, and liver carcinogenesis, and inhibited colon carcinogenesis. Excess of various forms of vitamin A enhanced or inhibited skin tumorigenesis, inhibited mammary carcinogenesis in rats (but not in mice), and carcinogenesis of the forestomach, liver, and urinary bladder (with one model, but not with another), or enhanced or did not influence lung carcinogenesis. Vitamin A analogs have enhanced or inhibited skin tumorigenesis, inhibited salivary gland, mammary, and urinary bladder carcinogenesis, enhanced tracheal and liver carcinogenesis, and either enhanced or inhibited pancreas carcinogenesis, depending upon the model employed. Although retinoids have been shown to inhibit carcinogenesis at many sites, numerous negative studies have been reported and some reports have indicated enhanced carcinogenesis. The most convincing evidence for the involvement of vitamin C in cancer prevention is the ability of ascorbic acid to prevent formation of nitrosamine and of other N-nitroso compounds. In addition vitamin C supplementation was shown to inhibit skin, nose, tracheal, lung, and kidney carcinogenesis, to either not influence or enhance skin, mammary gland, and colon carcinogenesis, and to enhance urinary bladder carcinogenesis, when given as sodium ascorbate, but not when given as ascorbic acid. Like vitamin C, vitamin E can inhibit nitrosation. Vitamin E was shown to inhibit skin, cheek pouch, and forestomach carcinogenesis, to enhance or inhibit colon carcinogenesis, and to have no effect on or to inhibit mammary gland carcinogenesis, depending upon the method of vitamin E administration or the level of dietary selenium or dietary fat. Selenium effects on carcinogenesis have been recently reviewed and the present discussi

Topics: Animals; Ascorbic Acid; Carcinogens; Cell Transformation, Neoplastic; Neoplasms, Experimental; Selenium; Vitamin A; Vitamin A Deficiency; Vitamin E

The behaviour of the C3H 10T1/2 C18 (10T1/2) cell line is reviewed in the context of its ability to reflect accurately events known to occur during carcinogenesis in vivo. It is concluded that, despite their embryonic fibroblastic origin and their infinite life-span in culture, 10T1/2 cells are capable of a wide range of responses to carcinogens and modulators of carcinogenesis that correspond closely to those observed in vivo, for the most part in epithelial tissues. For this and other reasons the 10T1/2 cell line has been widely employed in cancer research.

Topics: Animals; Antigens, Neoplasm; Ascorbic Acid; Biotransformation; Calcium; Carcinogens; Cell Adhesion; Cell Cycle; Cell Line; Cell Transformation, Neoplastic; Cells, Cultured; Culture Media; Dactinomycin; DNA; Glucocorticoids; Growth Substances; Mice; Neoplasms, Experimental; Oncogenes; Phorbol Esters; Protease Inhibitors; Retinoids; Saccharin; Structure-Activity Relationship

Management of oral leukoplakia-a potentially malignant disorder-is currently not evidence-based. Of the few randomized trials that have been reported, most have negative data. Therefore, a multi-centre, randomized, double-blind controlled trial (RCT) was undertaken to evaluate the use of low-dose beta-carotene combined with vitamin C supplements for the treatment and to prevent malignant transformation of oral leukoplakia. 46 Japanese participants with oral leukoplakia were allocated randomly either to an experimental arm (10 mg day(-1) of beta-carotene and 500 mg day(-1) of vitamin C) or placebo arm (50 mg day(-1) of vitamin C). Current or ex-smokers within 3 months of cessation were excluded. The supplements were continued over a period of 1 year. The primary endpoint was clinical remission at 1-year and the likelihood of malignant transformation during a 5-year follow-up period as a secondary endpoint. The overall clinical response rate in the experimental arm was 17.4% (4/23) and 4.3% (1/23) in the placebo arm (p = 0.346). During the median 60-month follow-up period, two subjects in the experimental arm and three in the control arm developed oral cancer. Under the intention-to-treat principle, relative risk by supplementing with beta-carotene and vitamin C was 0.77 (95%CI: 0.28-1.89) (p = 0.580) by the Cox proportional hazards model. No unfavorable side-effects were noted. Beta-carotene (10 mg day(-1) ) and vitamin C were neither effective for clinical remission, nor for protection against the development of cancer. Data from this RCT does not support the hypothesis that chemoprevention with this treatment is effective for oral leukoplakia.

Topics: Adult; Aged; Ascorbic Acid; beta Carotene; Cell Transformation, Neoplastic; Dietary Supplements; Disease Progression; Female; Follow-Up Studies; Humans; Leukoplakia, Oral; Male; Middle Aged; Risk Factors; Treatment Outcome

Gastric cancer is one of the most common malignancies worldwide. Histopathologic studies have identified a sequence of changes in the gastric mucosa that mark the slow progression from normal tissue to carcinoma. Epidemiologic evidence suggests that a diet rich in fresh fruit and vegetables could be a protective factor against this disease. This effect may be mediated through antioxidant vitamins.. A randomized, double-blind chemoprevention trial was conducted among 1980 subjects in Tachira State, Venezuela (whose population is at high risk for gastric cancer), to determine the effect of dietary supplementation with vitamin C, vitamin E, and beta-carotene on the progression and regression of precancerous gastric lesions. Subjects were randomly assigned to receive either a combination of vitamin C (750 mg/day), vitamin E (600 mg/day), and beta-carotene (18 mg/day) or placebo for 3 years. Changes in the gastric mucosa were determined by histologic diagnosis based on five biopsies taken from prespecified areas of the stomach at baseline and annually for 3 years. All biopsies were reviewed by a single expert pathologist. Progression rates (and regression rates) were calculated by comparing the first and last available gastroscopies for each subject and dividing the number of subjects whose diagnoses increased (decreased) in severity by the total follow-up time. Overall rate ratios were calculated by Poisson regression, controlling for baseline diagnosis. All statistical tests were two-sided.. Median plasma vitamin levels were increased in the treatment group between baseline and 1 year after randomization from 0.43 micromol/L (interquartile range [IQR] = 0.26-0.69) to 2.89 micromol/L (IQR = 1.76-4.22) for beta-carotene, from 26.7 micromol/L (IQR = 23.1-31.2) to 54.9 micromol/L (IQR = 42.8-67.6) for alpha-tocopherol, and from 47.70 micromol/L (IQR = 36.9-58.5) to 61.9 micromol/L (IQR = 52.2-72.7) for vitamin C. Overall progression rates per 100 person-years were 74.3 in the placebo group and 67.8 in the group randomly assigned to vitamins. Overall regression rates were 109.4 in the placebo group and 116.5 in the group randomly assigned to vitamins. There was no statistically significant difference in progression rate (rate ratio = 0.92, 95% confidence interval [CI] = 0.74 to 1.15) or regression rate (rate ratio = 1.09, 95% CI = 0.90 to 1.33) between vitamin and placebo groups.. Supplementation with antioxidant micronutrients is not an effective tool for gastric cancer control in this high-risk population. The results of this trial are consistent with previous findings on the lack of effect of nutritional supplementation on precancerous gastric lesions.

Topics: Adult; Aged; Antioxidants; Ascorbic Acid; beta Carotene; Cell Transformation, Neoplastic; Disease Progression; Double-Blind Method; Female; Gastric Mucosa; Gastroscopy; Helicobacter Infections; Helicobacter pylori; Humans; Logistic Models; Male; Middle Aged; Patient Selection; Precancerous Conditions; Risk Assessment; Sample Size; Smoking; Stomach Neoplasms; Treatment Failure; Venezuela; Vitamin E; Vitamins

Free radicals and reactive species produced in vivo can trigger cell damage and DNA modifications resulting in carcinogenesis. Dietary antioxidants trap these species limiting their damage. The present study evaluated the role of vitamins C and E in the prevention of potentially premalignant modifications to DNA in the human stomach by supplementing patients who, because of hypochlorhydria and possible depletion of gastric antioxidants, could be at increased risk of gastric cancer. Patients undergoing surveillance for Barrett's oesophagus (n 100), on long-term proton pump inhibitors were randomized into two groups: vitamin C (500 mg twice/d) and vitamin E (100 mg twice/d) for 12 weeks (the supplemented group) or placebo. Those attending for subsequent endoscopy had gastric juice, plasma and mucosal measurements of vitamin levels and markers of DNA damage. Seventy-two patients completed the study. Plasma ascorbic acid, total vitamin C and vitamin E were elevated in the supplemented group consistent with compliance. Gastric juice ascorbic acid and total vitamin C levels were raised significantly in the supplemented group (P=0.01) but supplementation had no effect on the mucosal level of this vitamin. However, gastric juice ascorbic acid and total vitamin C were within normal ranges in the unsupplemented group. Mucosal malondialdehyde, chemiluminescence and DNA damage levels in the comet assay were unaffected by vitamin supplementation. In conclusion, supplementation does not affect DNA damage in this group of patients. This is probably because long-term inhibition of the gastric proton pump alone does not affect gastric juice ascorbate and therefore does not increase the theoretical risk of gastric cancer because of antioxidant depletion.

Topics: Achlorhydria; Adult; Aged; Antacids; Antioxidants; Ascorbic Acid; Barrett Esophagus; Cell Transformation, Neoplastic; Dietary Supplements; DNA Damage; Female; Gastric Acidity Determination; Gastric Juice; Gastric Mucosa; Humans; Hydrogen-Ion Concentration; Male; Middle Aged; Precancerous Conditions; Proton Pump Inhibitors; Stomach Neoplasms; Vitamin E

Oxidative DNA damage may be important in mutagenic, carcinogenic, and aging processes. Although it is plausible that antioxidant vitamins may reduce oxidative DNA damage, evidence from human studies has been sparse and inconsistent. We determined the short-term effects of vitamin C (500 mg/day) and vitamin E (400 IU d-alpha-tocopheryl acetate/day) supplements on oxidative DNA damage in a double-masked, placebo-controlled, 2x2 factorial trial in 184 nonsmoking adults. Mean duration of supplementation was 2 months. Oxidative DNA damage was measured by 24-h urinary excretion of 8-hydroxy-2'-deoxyguanosine (8-OHdG). At baseline, urinary 8-OHdG (mean +/- SE; ng/mg creatinine) was associated with race (15.6 +/- 0.8 in African Americans versus 20.3 +/- 1.2 in Caucasians, P = 0.001), prior antioxidant supplement use (18.6 +/- 0.8 in users versus 13.8 +/- 1.5 in non-users, P = 0.007), and regular exercise (19.2 +/- 1.1 in exercisers versus 16.6 +/- 0.9 in non-exercisers, P = 0.04). Fruit and vegetable intake and serum ascorbic acid were inversely associated with urinary 8-OHdG (P-trend = 0.02 and 0.016, respectively). The benefits of fruit and vegetable intake became evident with the consumption being at least three servings/day. At the end of supplementation, change from baseline in urinary 8-OHdG (mean +/- SE; ng/mg creatinine) was -0.6 +/- 1.4 (P = 0.61), 0.6 +/- 1.1 (P = 0.59), 0.5 +/- 1.0 (P = 0.61), and 1.6 +/- 1.4 (P = 0.27) in the placebo, vitamin C alone, vitamin E alone, and combined vitamins C and E groups, respectively. In overall and subgroup analyses, there was no significant main effect or interaction effect of the supplements on urinary 8-OHdG. In conclusion, supplementation of diet with vitamin C (500 mg/day) and vitamin E (400 IU d-alpha-tocopheryl acetate/day) had no significant main effect or interaction effect on oxidative DNA damage as measured by urinary 8-OHdG in nonsmoking adults. However, several aspects of a healthy lifestyle were associated with lower oxidative DNA damage.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Aged; Antioxidants; Ascorbic Acid; Cell Transformation, Neoplastic; Chemoprevention; Deoxyguanosine; DNA Damage; Double-Blind Method; Female; Humans; Life Style; Male; Middle Aged; Oxidative Stress; Vitamin E

Previous research has identified a high risk of gastric carcinoma as well as a high prevalence of cancer precursor lesions in rural populations living in the province of Nariño, Colombia, in the Andes Mountains.. A randomized, controlled chemoprevention trial was conducted in subjects with confirmed histologic diagnoses of multifocal nonmetaplastic atrophy and/or intestinal metaplasia, two precancerous lesions. Individuals were assigned to receive anti-Helicobacter pylori triple therapy and/or dietary supplementation with ascorbic acid, beta-carotene, or their corresponding placebos. Gastric biopsy specimens taken at baseline were compared with those taken at 72 months. Relative risks of progression, no change, and regression from multifocal nonmetaplastic atrophy and intestinal metaplasia were analyzed with multivariate polytomous logistic regression models to estimate treatment effects. All statistical tests were two-sided.. All three basic interventions resulted in statistically significant increases in the rates of regression: Relative risks were 4.8 (95% confidence interval [CI] = 1.6-14.2) for anti-H. pylori treatment, 5. 1 (95% CI = 1.7-15.0) for beta-carotene treatment, and 5.0 (95% CI = 1.7-14.4) for ascorbic acid treatment in subjects with atrophy. Corresponding relative risks of regression in subjects with intestinal metaplasia were 3.1 (95% CI = 1.0-9.3), 3.4 (95% CI = 1.1-9.8), and 3.3 (95% CI = 1.1-9.5). Combinations of treatments did not statistically significantly increase the regression rates. Curing the H. pylori infection (which occurred in 74% of the treated subjects) produced a marked and statistically significant increase in the rate of regression of the precursor lesions (relative risks = 8.7 [95% CI = 2.7-28.2] for subjects with atrophy and 5.4 [95% CI = 1.7-17.6] for subjects with intestinal metaplasia).. In the very high-risk population studied, effective anti-H. pylori treatment and dietary supplementation with antioxidant micronutrients may interfere with the precancerous process, mostly by increasing the rate of regression of cancer precursor lesions, and may be an effective strategy to prevent gastric carcinoma.

Topics: Adult; Aged; Anti-Bacterial Agents; Antioxidants; Ascorbic Acid; beta Carotene; Biopsy; Cell Transformation, Neoplastic; Disease Progression; Drug Therapy, Combination; Female; Gastritis, Atrophic; Helicobacter Infections; Helicobacter pylori; Humans; Logistic Models; Male; Middle Aged; Precancerous Conditions; Remission, Spontaneous; Risk; Stomach; Stomach Neoplasms; Treatment Outcome

Mutagen sensitivity, as measured by an in vitro assay, has been described as a risk factor for the development of several tobacco-related epithelial cancers. In vitro studies have indicated that sensitivity to the clastogenic effects of bleomycin on chromosomes was reduced with the introduction of ascorbic acid in a dose-dependent relationship. We report the results of a randomized clinical trial to determine whether increasing levels of oral ascorbic acid could reduce the levels of mutagen sensitivity. For this study, we recruited 228 healthy smokers from 21 centers around the country through the Clinical Community Oncology Program. Each individual was randomly assigned to one of four daily regimens: placebo, 1 g of ascorbic acid, 2 g of ascorbic acid, or 4 g of ascorbic acid. Treatments were administered for 16 weeks. Assessment of mutagen sensitivity was made at baseline and at weeks 4, 16, and 20 (4 weeks after cessation of treatment). Serum ascorbic acid levels were measured at baseline and at weeks 4 and 16. Demographic and risk factor data were collected at baseline and at each-measurement point. Analyses measured the differences of mutagen sensitivity levels across the four treatment arms, as well as investigating the correlation between serum ascorbic acid level and mutagen sensitivity levels in individuals. We did not find a dose-response relationship between ascorbic acid intake and mutagen sensitivity. Additionally, we did not find an association between serum ascorbic acid levels and mutagen sensitivity.

Topics: Administration, Oral; Adult; Anticarcinogenic Agents; Ascorbic Acid; Bleomycin; Carcinogens; Cell Transformation, Neoplastic; Dose-Response Relationship, Drug; Female; Humans; In Vitro Techniques; Male; Middle Aged; Mutagenicity Tests; Smoking

A randomized nutrition intervention trial was conducted among 29,584 adult residents of Linxian, China, to examine the effects of vitamin/mineral supplementation on the occurrence of esophageal/gastric cardia cancer in this high-risk population. A fractional factorial study design allowed evaluations of four different combinations of nutrients: (A) retinol and zinc; (B) riboflavin and niacin; (C) vitamin C and molybdenum; and (D) beta-carotene, vitamin E, and selenium. During the 5.25-year intervention, significant reductions in total mortality, total cancer mortality, and stomach cancer mortality occurred among those receiving beta-carotene, vitamin E, and selenium. At the end of intervention, an endoscopic survey was carried out in a sample of subjects to see if the nutritional supplements had affected the prevalence of clinically silent precancerous lesions and early invasive cancers of the esophagus or stomach. Endoscopy was performed on 391 individuals from two study villages. The prevalences of esophageal and gastric dysplasia and cancer were compared by nutrient factor. Cancer or dysplasia was diagnosed in 15% of the participants. No statistically significant reductions in the prevalence of esophageal or gastric dysplasia or cancer were seen for any of the four vitamin/mineral combinations. The greatest reduction in risk (odds ratio, 0.38; P = 0.09) was seen for the effect of retinol and zinc on the prevalence of gastric cancer. Although no significant protective effects were seen in this endoscopic survey, there was a suggestion that supplementation with retinol and zinc may protect against the development of gastric neoplasia in this high-risk population. Additional studies with larger numbers of endpoints will be needed to further evaluate this possibility.

Topics: Adenocarcinoma; Adult; Aged; Ascorbic Acid; beta Carotene; Carotenoids; Cell Transformation, Neoplastic; China; Cross-Cultural Comparison; Cross-Sectional Studies; Double-Blind Method; Endoscopy, Gastrointestinal; Esophageal Neoplasms; Esophagus; Female; Gastric Mucosa; Humans; Incidence; Male; Middle Aged; Minerals; Molybdenum; Niacin; Precancerous Conditions; Riboflavin; Rural Population; Selenium; Stomach Neoplasms; Survival Rate; Vitamin A; Vitamin E; Vitamins; Zinc

Linxian, China has some of the highest rates of esophageal/gastric cardia cancer in the world, and epidemiological evidence suggests that chronically low intake of micronutrients may contribute to these high cancer rates. To examine whether supplementation with multiple vitamins and minerals can affect the occurrence of esophageal/gastric cardia cancer in this population, a two-arm randomized nutrition intervention trial was conducted among 3318 Linxian residents with cytological evidence of esophageal dysplasia. During the 6-year intervention, esophageal/gastric cardia cancer mortality was 8% lower among those receiving the active supplements. After 30 and 72 months of intervention, endoscopic surveys were carried out to see if the nutritional supplements had affected the prevalence of clinically silent precancerous lesions and early invasive cancers of the esophagus and stomach. In the first survey, in 1987, 833 subjects were endoscoped; in the second survey, in 1991, 396 subjects were examined. The histological diagnoses from each survey were compared by treatment group. Cancer or dysplasia was diagnosed in 28% of the subjects endoscoped in 1987 and 24% of those examined in 1991. The odds ratio for subjects in the treatment group (versus those in the placebo group) having esophageal or gastric dysplasia or cancer was 0.84 (95% confidence interval, 0.61-1.15) in 1987 and 0.86 (0.54-1.38) in 1991. Although modest protective effects on worst overall diagnosis were seen in the supplemented group in both surveys, none of the results was statistically significant, and the findings must be considered inconclusive.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenocarcinoma; Adult; Aged; Ascorbic Acid; beta Carotene; Carotenoids; Cell Transformation, Neoplastic; Cross-Cultural Comparison; Double-Blind Method; Endoscopy, Gastrointestinal; Esophageal Neoplasms; Esophagus; Female; Follow-Up Studies; Gastric Mucosa; Humans; Male; Middle Aged; Minerals; Molybdenum; Niacin; Precancerous Conditions; Riboflavin; Rural Population; Selenium; Stomach Neoplasms; Survival Rate; Vitamin A; Vitamin E; Vitamins; Zinc

Quercetin (Que) is an abundant flavonoid in the human diet and high-concentration food supplement with reported pro- and anti-carcinogenic activities. Topoisomerase II (TopoII) inhibition and subsequent DNA damage induction by Que was implicated in the mixed lineage leukemia gene (MLL) rearrangements that can induce infant and adult leukemias. This notion raised concerns regarding possible genotoxicities of Que in hematopoietic stem and progenitor cells (HSPCs). However, molecular targets mediating Que effects on DNA repair relevant to MLL translocations have not been defined. In this study we describe novel and potentially genotoxic Que activities in suppressing non-homologous end joining and homologous recombination pathways downstream of MLL cleavage. Using pharmacological dissection of DNA-PK, ATM and PI3K signalling we defined PI3K inhibition by Que with a concomitant decrease in the abundance of key DNA repair genes to be responsible for DNA repair inhibition. Evidence for the downstream TopoII-independent mutagenic potential of Que was obtained by documenting further increased frequencies of MLL rearrangements in human HSPCs concomitantly treated with Etoposide and Que versus single treatments. Importantly, by engaging a tissue engineered placental barrier, we have established the extent of Que transplacental transfer and hence provided the evidence for Que reaching fetal HSPCs. Thus, Que exhibits genotoxic effects in human HSPCs via different mechanisms when applied continuously and at high concentrations. In light of the demonstrated Que transfer to the fetal compartment our findings are key to understanding the mechanisms underlying infant leukemia and provide molecular markers for the development of safety values.

Topics: Adult; Ascorbic Acid; Cell Culture Techniques; Cell Transformation, Neoplastic; Cells, Cultured; Colony-Forming Units Assay; DNA Damage; DNA Repair; DNA Topoisomerases, Type II; Dose-Response Relationship, Drug; Etoposide; Female; Genistein; Hematopoietic Stem Cells; Histone-Lysine N-Methyltransferase; Histones; Humans; Infant; Leukemia; Maternal-Fetal Exchange; Myeloid-Lymphoid Leukemia Protein; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Pregnancy; Quercetin; Signal Transduction; Topoisomerase II Inhibitors

Nitrate and nitrite are precursors of endogenously formed N-nitroso compounds (NOC), known animal carcinogens. Nitrosation reactions forming NOCs can be inhibited by vitamin C and other antioxidants. We prospectively investigated the association between dietary nitrate and nitrite intake and risk of colorectal cancer in the Shanghai Women's Health Study, a cohort of 73,118 women ages 40-70 residing in Shanghai. We evaluated effect modification by factors that affect endogenous formation of NOCs: vitamin C (at or above/below median) and red meat intake (at or above/below median). Nitrate, nitrite and other dietary intakes were estimated from a 77-item food frequency questionnaire administered at baseline. Over a mean of 11 years of follow-up, we identified 619 colorectal cancer cases (n = 383, colon; n = 236, rectum). Hazard ratios (HR) and 95% confidence intervals (CI) were estimated using Cox proportional hazard regression. Overall, nitrate intake was not associated with colorectal cancer risk (HR = 1.08; 95% CI: 0.73-1.59). However, among women with vitamin C intake below the median (83.9 mg day(-1) ) and hence higher potential exposure to NOCs, risk of colorectal cancer increased with increasing quintiles of nitrate intake (highest vs. lowest quintile HR = 2.45; 95% CI: 1.15-5.18; p trend = 0.02). There was no association among women with higher vitamin C intake. We found no association between nitrite intake and risk of colorectal cancer overall or by intake level of vitamin C. Our findings suggest that high dietary nitrate intake among subgroups expected to have higher exposure to endogenously formed NOCs increases risk of colorectal cancer.

Topics: Adult; Aged; Ascorbic Acid; Cell Transformation, Neoplastic; China; Colorectal Neoplasms; Diet; Feeding Behavior; Female; Humans; Meat; Middle Aged; Nitrates; Nitrites; Nitroso Compounds; Prospective Studies; Risk; Surveys and Questionnaires; Women's Health

MicroRNAs (miRNA) are small non-coding RNAs that regulate the expression of approximately 60% of all human genes and play important roles in disease processes. Recent studies have demonstrated a link between dysregulated expression of miRNAs and breast carcinogenesis. Long-term estrogen exposure is implicated in development of human breast cancers, yet underlying mechanisms remain elusive. We have recently demonstrated that antioxidant vitamin C (vit C) prevents estrogen-induced breast tumor development. In this study, we investigated the role of vit C in the regulation of microRNA-93 (miR-93) and its target gene(s) in a rat model of mammary carcinogenesis. Female August Copenhagen Irish (ACI) rats were treated with vit C in the presence or absence of 17β-estradiol (E2) for 8 months. We demonstrate an increased expression of the miR-93 in E2-treated mammary tissues and in human breast cell lines and vit C treatment reverted E2-mediated increase in miR-93 levels. MiRNA target prediction programs suggest one of the target genes of miR-93 to be nuclear factor erythroid 2-related factor 2 (NRF2). In contrast with miR-93 expression, NRF2 protein expression was significantly decreased in E2-treated mammary tissues, mammary tumors, and in breast cancer cell lines, and its expression was significantly increased after vit C treatment. Ectopic expression of miR-93 decreased protein expression of NRF2 and NRF2-regulated genes. Furthermore, miR-93 decreased apoptosis, increased colony formation, mammosphere formation, cell migration and DNA damage in breast epithelial cells, whereas silencing of miR-93 in these cells inhibited these carcinogenic processes. Taken together, our findings suggest an oncogenic potential of miR-93 during E2-induced breast carcinogenesis.

Topics: Animals; Apoptosis; Ascorbic Acid; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Cell Transformation, Neoplastic; DNA Damage; Estradiol; Estrogens; Female; Humans; MicroRNAs; NF-E2-Related Factor 2; Rats; Rats, Inbred ACI; Rats, Sprague-Dawley; Up-Regulation

Ascorbic acid (Vitamin C) administration has been used to prevent infectious diseases in public or as a therapeutic agent by the physicians in treatment of several diseases. Ascorbic acid is also involved in immune cell functions and immune responses, although the mechanisms by which it exerts effects on immune cells against cancer cells are not fully understood at the normal plasma level. In this study, we used the mice lacking l-gulono-γ-lactone oxidase (Gulo), the enzyme required for the biosynthesis of ascorbic acid, to characterize the effects of ascorbic acid on NK cell cytotoxicity against ovarian cancer cells, MOSECs (murine ovarian surface epithelial cells). Gulo(-/-) mice depleted of ascorbic acid survived for a shorter time than the normal control or Gulo(-/-) mice supplemented with ascorbic acid after tumor challenge regardless of treatment with IL-2. CD69 and NKG2D expression was clearly reduced in NK cells isolated from mice depleted of ascorbic acid as compared to that in the normal control and the mice supplemented with ascorbic acid. We also observed that IFN-γ secretion by NK cells isolated from Gulo(-/-) mice depleted of ascorbic acid was decreased after NK cells were co-cultured with MOSECs. Furthermore, the mRNA expression of perforin and granzyme B genes was also significantly decreased in NK cells isolated from mice depleted of ascorbic acid. Taken together, our results suggest that ascorbic acid at the normal plasma concentration has an essential role in maintaining the NK cytotoxicity against cancer cells.

Topics: Animals; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Antioxidants; Apoptosis; Ascorbic Acid; Cell Line, Tumor; Cell Transformation, Neoplastic; Cytotoxicity, Immunologic; Disease Models, Animal; Female; Granzymes; Killer Cells, Natural; Lectins, C-Type; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Knockout; NK Cell Lectin-Like Receptor Subfamily K; Ovarian Neoplasms; Perforin

The mechanisms underlying the pathogenesis of estrogen-induced breast carcinogenesis remain unclear. The present study investigated the roles of estrogen metabolism and oxidative stress in estrogen-mediated mammary carcinogenesis in vivo. Female August Copenhagen Irish (ACI) rats were treated with 17beta-estradiol (E(2)), the antioxidant vitamin C, the estrogen metabolic inhibitor alpha-naphthoflavone (ANF), or cotreated with E(2) + vitamin C or E(2) + ANF for up to 8 months. E(2) (3 mg) was administered as an subcutaneous implant, ANF was given via diet (0.2%) and vitamin C (1%) was added to drinking water. At necropsy, breast tumor incidence in the E(2), E(2) + vitamin C and E(2) + ANF groups was 82, 29 and 0%, respectively. Vitamin C and ANF attenuated E(2)-induced alterations in oxidative stress markers in breast tissue, including 8-iso-prostane F(2alpha) formation and changes in the activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase. Quantification of 2-hydroxyestradiol (2-OHE(2)) and 4-hydroxyestradiol (4-OHE(2)) formation in breast tissue confirmed that ANF inhibited 4-hydroxylation of E(2) and decreased formation of the highly carcinogenic 4-OHE(2). These results demonstrate that antioxidant vitamin C reduces the incidence of estrogen-induced mammary tumors, increases tumor latency and decreases oxidative stress in vivo. Further, our data indicate that ANF completely abrogates breast cancer development in ACI rats. The present study is the first to demonstrate the inhibition of breast carcinogenesis by antioxidant vitamin C or the estrogen metabolic inhibitor ANF in an animal model of estrogen-induced mammary carcinogenesis. Taken together, these results suggest that E(2) metabolism and oxidant stress are critically involved in estrogen-induced breast carcinogenesis.

Topics: Animals; Antioxidants; Ascorbic Acid; Benzoflavones; Cell Transformation, Neoplastic; Dinoprost; Estradiol; Estrogens, Catechol; Female; Mammary Neoplasms, Experimental; Neoplasms, Hormone-Dependent; Oxidative Stress; Rats; Rats, Inbred ACI

The incidence of esophageal adenocarcinoma (EA) and its precursor condition, Barrett's esophagus, has risen rapidly in the United States for reasons that are not fully understood. Therefore, we evaluated the association between use of supplemental vitamins and minerals and risk of neoplastic progression of Barrett's esophagus and EA. The Seattle Barrett's Esophagus Program is a prospective study based on 339 men and women with histologically confirmed Barrett's esophagus. Participants underwent baseline and periodic follow-up exams, which included endoscopy and self-administered questionnaires on diet, supplement use, and lifestyle characteristics. Use of multivitamins and 4 individual supplements was calculated using time-weighted averages of reported use over the observational period. Cox proportional-hazards models were used to calculate hazard ratios (HR) for each endpoint: EA, tetraploidy, and aneuploidy. During a mean follow-up of 5 yr, there were 37 cases of EA, 42 cases of tetraploidy, and 34 cases of aneuploidy. After controlling for multiple covariates including diet, nonsteroidal anti-inflammatory drug use, obesity, and smoking, participants who took 1 or more multivitamin pills/day had a significantly decreased risk of tetraploidy [HR = 0.19; 95% confidence interval (CI) = 0.08-0.47) and EA (HR = 0.38; 95% CI = 0.15-0.99] compared to those not taking multivitamins. Significant inverse associations were also observed between risk of EA and supplemental vitamin C (> or = 250 mg vs. none: HR = 0.25; 95% CI = 0.11-0.58) and vitamin E (> or = 180 mg vs. none: HR = 0.25; 95% CI = 0.10-0.60). In this cohort study, use of multivitamins and single antioxidant supplements was associated with a significantly reduced risk of EA and markers of neoplastic progression among individuals with Barrett's esophagus.

Topics: Adenocarcinoma; Adult; Aged; Aneuploidy; Ascorbic Acid; Barrett Esophagus; Cell Transformation, Neoplastic; Chemoprevention; Cohort Studies; Dietary Supplements; Disease Progression; Drug Therapy, Combination; Endoscopy; Esophageal Neoplasms; Female; Flow Cytometry; Follow-Up Studies; Humans; Male; Middle Aged; Proportional Hazards Models; Prospective Studies; Risk Factors; Vitamin E; Vitamins

The luminal microenvironment including acid and nitric oxide (NO) has been implicated in Barrett's esophagus carcinogenesis. We investigated the ability of acid and NO to induce DNA damage in esophageal cells.. Transformed and primary Barrett's esophagus and adenocarcinoma cells were exposed to either acid, (pH 3.5), +/- antioxidant or NO from a donor or generated by acidification of nitrite in the presence of ascorbate +/- NO scavenger. Phosphorylation of histone H2AX and the neutral comet assay were used to detect DNA double-strand breaks (DSBs). Intracellular levels of reactive oxygen species and NO were detected with fluorescent dyes. Mitochondrial viability was measured with a rhodamine dye. Long-term survival was assessed by clonogenic assay.. Exposure to acid (pH 3.5) for > or =15 minutes induced DSBs in all cell lines (P < .05). There was a concomitant increase in intracellular reactive oxygen species in the absence of mitochondrial damage, and pretreatment with antioxidants inhibited DNA damage. Exposure to physiologic concentrations of NO produced from the NO donor or acidification of salivary nitrite induced DSBs in a dose- (>25 micromol/L) and cell-dependent manner (adenocarcinoma >Barrett's esophagus, P < .05). This occurred preferentially in S-phase cells consistent with stalled replication forks and was blocked with a NO scavenger. NO also induced DSBs in primary Barrett's esophagus cells treated ex vivo. Cells were able to survive when exposed to acid and NO.. Both acid and NO have the potential to generate DSBs in the esophagus and via distinct mechanisms.

Topics: Adenocarcinoma; Antioxidants; Ascorbic Acid; Barrett Esophagus; Cell Line, Tumor; Cell Survival; Cell Transformation, Neoplastic; Comet Assay; DNA Breaks, Single-Stranded; Dose-Response Relationship, Drug; Esophageal Neoplasms; Gastric Acid; Histones; Humans; Hydrazines; Hydrogen-Ion Concentration; Mitochondria; Nitric Oxide; Nitric Oxide Donors; Phosphorylation; Reactive Oxygen Species; S Phase; Sodium Nitrite; Time Factors

Selenium, in the form of 1,4-phenylenebis(methylene)selenocyanate (p-XSC) but not Se-enriched yeast (Se-yeast), was highly effective at inhibiting lung tumors induced by the tobacco specific nitrosamine (TSNA) 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in A/J mice and at reducing NNK-induced DNA methylation and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels in the lung. Our goal was to determine if p-XSC but not Se-yeast is effective at inducing levels of glutathione (GSH)-related antioxidants and reducing markers of GSH oxidation in the NNK-induced lung tumor model. In the first bioassay, 6-week-old mice were fed either control or experimental diets (containing 10 ppm as selenium from p-XSC or Se-yeast) and, beginning at 8 weeks of age, received NNK (3 micromol) by gavage once weekly for 8 weeks. After 18 weeks, p-XSC significantly reduced NNK-induced tumor burden by 74% (10.4 +/- 6.0 versus 2.7 +/- 1.5 tumors/mouse, P < 0.001) and tumor incidence from 96% to 68% (P < 0.01), whereas, Se-yeast had no effect. Lung GSH levels were unchanged by either NNK or Se-yeast, but were increased 70% in mice treated with both NNK and p-XSC (P < 0.01) and 41% in mice treated with p-XSC alone. In the second bioassay, the time course of effects of p-XSC was examined. As early as one week after initiation of p-XSC feeding lung and blood selenium levels were increased nearly six- and two-fold, respectively. Increases of 120% for GSH and 65% for Cys were observed in p-XSC groups compared to controls within one week after initiation of p-XSC feeding (P < 0.01). The levels of protein-bound:free GSH ratios and Cys ratios were significantly decreased in p-XSC-treated mice, regardless of NNK status, suggesting a decrease in the levels of oxidative stress. Altogether, these results indicate that p-XSC is a potent inducer of GSH and related thiol antioxidants in the lung leading to decreased levels of oxidative stress and suggest that p-XSC inhibits tumor formation, in part, by protecting against oxidative damage.

Topics: Animal Feed; Animals; Antioxidants; Ascorbic Acid; Cell Transformation, Neoplastic; Cysteine; Disulfides; Female; Glutathione; Lung Neoplasms; Mice; Molecular Structure; Nitrosamines; Organoselenium Compounds; Selenium; Sulfhydryl Compounds; Time Factors; Yeasts

To evaluate the potential role of oxidative stress in the evolution of cervical cancer, including its pre-malignant states.. Erythrocytes thiobarbituric acid reactive substances (TBARS) levels, plasma vitamin C and thiol content and total blood delta-ALA-D levels were estimated in 46 untreated cervical cancer and pre-malignant patients and in 46 age-sex-matched controls.. Erythrocytes from patients, regardless of disease state, pre-malignant (low squamous intraepithelial lesion--LSIL and high squamous intraepithelial lesion--HSIL) or cancer, showed a significant 2-3 times increase in TBARS levels (P<0.01). Plasma vitamin C was lower in the carcinoma group (P<0.01). The reactivation index of delta-aminolevulinate dehydratase (delta-ALA-D) was higher in the patient group, when compared to control (P<0.01).. LSIL, HSIL or cervical cancer can be associated with changes in 3 indicators of oxidative stress: increase in erythrocyte TBARS, ALA-D reactivation index and a decrease in vitamin C content, that may play an important role in carcinogenesis.

Topics: Adult; Ascorbic Acid; Cell Transformation, Neoplastic; Erythrocytes; Female; Humans; Middle Aged; Oxidative Stress; Porphobilinogen Synthase; Sulfhydryl Compounds; Thiobarbituric Acid Reactive Substances; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Syrian hamster embryo (SHE) cell transformation has been used for many years to study chemical carcinogenesis in vitro. It has been shown that this assay is probably the most predictive short-term test system for identifying rodent carcinogens. Although most of the operational difficulties encountered in the early stage of application of this assay have been overcome by culturing the SHE cells under slightly acidic conditions (pH 6.7), a relatively low level of induction of morphological transformation (MT) by known carcinogens still occurs for many cell isolates. In order to improve the response of this assay system to known carcinogens, the effect of incubation time of target SHE cells on the frequency of morphological transformation induced by benzo(a)pyrene (BaP) was investigated. It was shown that the morphological transformation frequency induced by BaP increased significantly (1.4-2.5-fold) when the incubation time of target cells was reduced from the usual 24h to less than 6h prior to seeding onto feeder layers. This improvement in sensitivity was consistent for different cell isolates. In addition, the enhanced response appeared to be a property of carcinogens because treatment with two non-carcinogens, l-ascorbic acid and 4-nitro-o-phenylenediamine, did not induce significant increases in the transformation frequency under the shortened incubation period for target cells. These results suggest that the response of the SHE cell transformation assay may be improved by optimizing the incubation time of the target SHE cells. In addition, the results of the present study provide further evidence to support the idea that morphological transformation of SHE cells results from a block of cellular differentiation of stem or stem-like cells.

Topics: Animals; Antioxidants; Ascorbic Acid; Benzo(a)pyrene; Carcinogenicity Tests; Carcinogens; Cell Survival; Cell Transformation, Neoplastic; Cells, Cultured; Cellular Senescence; Cricetinae; Mesocricetus; Phenylenediamines; Time Factors

Telomerase activation can be considered as a critical step in cell immortalization. The enzyme elongates or maintains telomere length by adding to its end tandem TTAGGG repeats by using its endogenous RNA template. Telomerase is not detectable in most somatic cells but is upregulated in germ line cells and in 85-90% of human cancers, which suggests important role of telomerase in neoplastic transformation. Consequently, telomerase has been proposed as a potentially highly selective target for the development of antiproliferative agents. Platinum complexes are widely administrated in cancer therapy. A conjugate of selenite with diammineplatinum [(NH(3))(2)Pt(SeO(3))(2)] is a novel potential anticancer drug. Using alkaline single cell gel electrophoresis (comet assay), we showed that the drug at 5-30 microM induced concentration-dependent damage to DNA of endometrial cancer cells derived from tumor samples. Sodium ascorbate at 10 and 50 microM reduced the extent of the DNA damage evoked by the drug. (NH(3))(2)Pt(SeO(3)) reduced telomerase activity in the cells in a concentration-dependent manner as measured by using the telomere repeat amplification protocol (TRAP) assay. This effect was independent of sodium ascorbate. Therefore, mutagenic effects of the conjugate can be reduced by well-recognized antimutagen, sodium ascorbate, but it can still retain ability to affect neoplastic transformation. The results obtained indicate that (NH(3))(2)Pt(SeO(3)) may specifically inhibit telomerase activity in endometrial cancer cells.

Topics: Antineoplastic Agents; Antioxidants; Ascorbic Acid; Cell Survival; Cell Transformation, Neoplastic; Cisplatin; Comet Assay; DNA Damage; Dose-Response Relationship, Drug; Endometrial Neoplasms; Female; Humans; Polymerase Chain Reaction; Selenium Compounds; Telomerase; Telomere

Catechol estrogens are considered critical intermediates in estrogen-induced carcinogenesis. We demonstrated previously that 17beta-estradiol (E(2)), estrone (E(1)) and four of their catechol estrogens, 2- and 4-hydroxyestradiols (2- and 4-OHE(2)), and 2- and 4-hydroxyestrones (2- and 4-OHE(1)) induce morphological transformation in Syrian hamster embryo (SHE) fibroblasts, and the transforming abilities vary as follows: 4-OHE(1) > 2-OHE(1) > 4-OHE(2) > 2-OHE(2) vertical line E(2), E(1). To examine the involvement of catechol estrogens in the initiation of hormonal carcinogenesis, we studied the ability of E(2), E(1) and their catechol estrogens to induce DNA adducts in SHE cells by using a (32)P-post-labeling assay. DNA adducts were detected in cells treated with each of all the catechol estrogens at concentrations of 10 microg/ml for 1 h and more. 2- or 4-OHE(2) formed a single DNA adduct, which was chromatographically distinct from each other. In contrast, 2- or 4-OHE(1) produced one major and one minor adduct, and the two adducts formed by each catechol estrogen exhibited identical mobilities on the chromatograms. Neither E(2) nor E(1) at concentrations up to 30 microg/ml induced DNA adducts. The abilities of the estrogens to induce DNA adducts were ranked as follows: 4-OHE(1) > 2-OHE(1) > 4-OHE(2) > 2-OHE(2) > > E(2), E(1), which corresponds well to the transforming and carcinogenic abilities of the estrogens. In addition, the level of DNA adducts induced by the catechol estrogens was markedly decreased by co-treatment of cells with the antioxidant L-ascorbic acid. The results indicate the possible involvement of oxidative metabolites of catechol estrogens of E(2) and E(1) in the initiation of endogenous estrogen-induced carcinogenesis.

Topics: Animals; Antioxidants; Ascorbic Acid; Cell Survival; Cell Transformation, Neoplastic; Cricetinae; DNA Adducts; Estradiol; Estrogens, Catechol; Fibroblasts; Hydroxyestrones; Mesocricetus

In order to study the protection of beta-carotene and vitamin C against the transformation of human lung fibroblasts (HLF) cell induced by nickel sesquioxide, HLF cells were treated repeatedly by different concentrations of Ni2O3 in vitro. beta-carotene (5.4 mg/L) and vitamin C(1.8 mg/L) were added into the media containing Ni2O3(1.0 mg/L) respectively. The identification of malignancy of the transformation of HLF cell was carried out by the tests of ConA and the growth on semisolid agar culture. The results showed that Ni2O3 could induce the malignant transformation of HLF cell. The transformed cell proliferated rapidly. The transformed colonies exhibited in extensively random orientation and the cells were crossingover. The frequency of transformation showed a dose-response relation at the experimental concentrations. The transformed cell could be agglutinated by lower concentration of ConA and could grow in semisolid agar. The frequencies of transformation of HLF cell exposed to beta-carotene and vitamin C were decreased significantly. The cells could not be agglutinated by ConA and not grow in semisolid agar. It was concluded that Ni2O3 could induce strongly the malignant transformation of HLF cell and might be carcinogenic to human. The protection of beta-carotene and vitamin C on the transformation of HLF cell induced by Ni2O3 was observed. The increase of foods riched in beta-carotene and vitamin C was suggested for workers exposed to nickel.

Topics: Antioxidants; Ascorbic Acid; beta Carotene; Cell Transformation, Neoplastic; Cells, Cultured; Fibroblasts; Lung; Nickel

Both ascorbic acid (AsA, vitamin C) and N-acetylcysteine (NAC), a precursor and analogue of glutathione, possess a broad array of biological properties underlying their protective role in a variety of pathophysiological conditions. However, under certain circumstances, AsA behaves as a pro-oxidant rather than an anti-oxidant and produces adverse effects. This prompted us to evaluate whether NAC could interact with AsA in preventing mutation and cancer. AsA significantly increased spontaneous revertants in the Salmonella typhimurium strains TA102 and TA104, which are sensitive to oxidative mutagens. In contrast, NAC lowered the spontaneous background in TA104 and neutralized the negative effects of AsA. Moreover, NAC and AsA showed additive effects in reducing chromium(VI) and in reverting its mutagenicity. A single i.p. injection of urethane (1 g/kg body weight) to 120 A/J mice resulted, after 4 months, in the formation of a total of 1,532 lung tumors, 425 in the 30 mice treated with the carcinogen only, 404 in those treated with urethane plus AsA, 365 in those treated with urethane plus NAC and 338 in those treated with urethane plus the combination of AsA and NAC (both given daily with drinking water at the dose of 1 g/kg body weight). Compared to positive controls, tumor multiplicity was poorly affected by AsA, whereas it was significantly decreased by NAC and even more so by its combination with AsA. The overall volumes of lung tumors in the 4 groups were 107.5, 89.3, 61.3 and 49.7 mm(3), respectively. Tumor sizes were slightly but significantly decreased in mice treated with AsA and more so in those treated with NAC and NAC plus AsA, their combination being significantly more effective than each individually. All protective effects elicited by combining the 2 drugs were additive. Therefore, NAC prevents the adverse effects of AsA on spontaneous mutagenicity; at the same time, this thiol behaves in an additive fashion with AsA, inhibiting the mutagenicity of chromium(VI) and the lung tumorigenicity of urethane in mice. These findings suggest that NAC and AsA could conveniently be combined in cancer chemoprevention and other pharmacological interventions.

Topics: Acetylcysteine; Animals; Ascorbic Acid; Carcinogenicity Tests; Cell Transformation, Neoplastic; Chromium; Drug Stability; Female; Lung Neoplasms; Mice; Mutagenesis; Urethane

The anti-transforming effects of sodium ascorbate and its stable derivatives were examined in the two-stage transformation assay. When BALB/c 3T3 cells were treated with 0.2 microg/ml 20-methylcholanthrene as an initiator, and 100 ng/ml 12-O-tetradecanoylphorbol-13-acetate as a promoter, the addition at the promotion stage of L-ascorbic acid-2-phosphate ester magnesium (APM) was most marked in the inhibition of transformation. The inhibitory effects of sodium ascorbate and ascorbic acid-2-glucoside (AG) were comparable, but weaker than those of APM; L (+)-ascorbic acid-2-sulfate ester disodium 2H(2)O showed little effect. When phorbol 12, 13-didecanoate or tumor necrosis factor alpha (TNF-alpha) were used as promoters, APM also effectively suppressed transformation.

Topics: 3T3 Cells; Animals; Ascorbic Acid; Cell Transformation, Neoplastic; Dose-Response Relationship, Drug; Methylcholanthrene; Mice; Mice, Inbred BALB C; Tetradecanoylphorbol Acetate; Time Factors; Tumor Necrosis Factor-alpha

To examine the effects of ascorbic acid (AA) on hepatoma.. Choosing an all-trans tretinoin (Tre) as a positive control, cell growth, and cell redifferentiation tests by cell surface charges, biochemical changes, and cell growth in soft agar were measured.. After being treated with AA 6 mmol.L-1, the growth curve and mitotic index of human hepatoma cells decreased remarkably, the cellular growth inhibitory rate amounted to 58.9%. The indices related with cell malignancy alleviated, such as cell surface charge obviously decreased, the electrophoresis rate dropped from 1.64 microns.s-1.V-1.cm-1 to 0.93, the average value of alpha-fetoprotein (alpha-FP) content decreased from 302 micrograms.g-1(protein) to 90, and gamma-glytamyl-transpeptidase (gamma-GT) activity from 0.81 U.g-1(protein) to 0.16. The index related with cell differentiation increased, such as the average level of tyrosine-alpha-ketoglutarate transminase activity increased from 10.3 micromol.g-1(protein) to 41.2, and the colonogenic potential decreased 94.4%.. AA can inhibit human hepatoma cells proliferation, induce redifferentiation, and reverse its malignant phenotypic characteristics.

Topics: alpha-Fetoproteins; Antineoplastic Agents; Antioxidants; Ascorbic Acid; Carcinoma, Hepatocellular; Cell Division; Cell Survival; Cell Transformation, Neoplastic; gamma-Glutamyltransferase; Humans; Liver Neoplasms; Mitotic Index; Tumor Cells, Cultured

Using electronic spin resonance (ESR), we found a new type of radical with a long life-time in cells (T1/2>20 h) and which may play a more important role in the induction of mutation and transformation than either the active, short-lived, H, or OH radicals. When cells were treated with dimethyl sulfoxide (DMSO) and l-ascorbic acid (AsA) just before irradiation, the short-lived radicals were well-scavenged. On the other hand, if cells were treated with the scavengers 20 min after irradiation, then AsA scavenged the long-lived radicals, but DMSO did not. AsA treatment 20 min after the start of irradiation drastically reduced both the frequencies of mutation at the hypoxanthine guanine phosphoribosyl transferase (HGPRT) locus in human cells and morphological transformations in mouse m5S cells, but DMSO treatment did not. In addition, AsA treatment 20 h after irradiation also reduced the mutation frequency in human cells. These results suggested that mutations and morphological transformation are probably caused by the presence of long-lived radicals in the cells, rather than by short lived radicals, and that AsA reacts efficiently with long-lived radicals, resulting in a decrease of the mutations and transformations induced.

Topics: Animals; Ascorbic Acid; Cell Line; Cell Transformation, Neoplastic; Chromosome Aberrations; Dimethyl Sulfoxide; Electron Spin Resonance Spectroscopy; Free Radical Scavengers; Free Radicals; Half-Life; Humans; Hypoxanthine Phosphoribosyltransferase; Mice; Mutation; Radiation Tolerance

The effects of a synthetic phenolic antioxidant, 1-O-hexyl-2,3,5-trimethylhydroquinone (HTHQ), two novel synthetic ascorbic acid derivatives, 3-O-ethyl ascorbic acid (EAsA) and 3-O-dodecylcarbomethylascorbic acid (DAsA), and phenylethyl isothiocyanate (PEITC) on 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary carcinogenesis were examined in female Sprague-Dawley rats. Groups of 20, 7 week-old rats received an intra-gastric dose (50 mg/kg, b.w.) of DMBA, and starting one week thereafter received powdered diet containing 1.0% HTHQ, 1.0% EAsA, 1.0% DAsA, 0.1% PEITC or a basal diet alone for 35 weeks. Although the final incidences of mammary adenocarcinomas did not significantly differ among the DMBA-treated groups, multiplicities were significantly lowered in the EAsA (1.6+/-1.6 per rat, P < 0.01) and HTHQ (2.6+/-1.9, P < 0.05) animals as compared with the basal diet case (4.1+/-2.9). The average carcinoma volumes were also significantly smaller in rats given EAsA (2.1+/-3.8 cm3, P < 0.05), DAsA (2.5+/-5.3, P < 0.05) or PEITC (2.4+/-5.9, P < 0.05) than in those receiving DMBA alone (4.9+/-9.2). The results indicate that HTHQ, EAsA and PEITC all exert chemopreventive influence on the promotion/progression stage of DMBA-induced rat mammary carcinogenesis, with EAsA being particularly effective. To our knowledge this is the first documented example of an ascorbic acid derivative possessing chemopreventive potential against mammary cancer in vivo.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Adenocarcinoma; Animals; Anticarcinogenic Agents; Antioxidants; Ascorbic Acid; Carcinogens; Cell Transformation, Neoplastic; Chemoprevention; Diet; Female; Hydroquinones; Isothiocyanates; Mammary Neoplasms, Experimental; Rats; Rats, Sprague-Dawley

We have previously shown that trivalent chromium, and hexavalent chromium in the presence of one of its primary in vivo reductants, ascorbate, can bind to DNA and form interstrand crosslinks capable of obstructing replication. This effect was demonstrated in vitro by using Sequenase Version 2.0 T7 DNA polymerase; its parent enzyme, the unmodified T7 DNA polymerase; and Escherichia coli polymerase I large (Klenow) fragment; and it was demonstrated ex vivo by using Taq polymerase and DNA from chromium-treated human lung cells as template. This study was performed to determine whether DNA-bound chromium affects mammalian DNA polymerases in the same manner. Two mammalian enzymes, DNA polymerase alpha and DNA polymerase beta, were used. DNA polymerase alpha is a processive enzyme believed to be the primary lagging-stand synthetase, whereas DNA polymerase beta is a non-processive enzyme believed to function in DNA repair by filling single stranded gaps one base at a time. DNA polymerase arrest assays were performed with each of these enzymes to replicate DNA with toxicologically relevant levels of chromium adducts produced by either trivalent chromium or hexavalent chromium and ascorbate. Both enzymes responded to chromium-DNA damage by arresting replication, and the arrests increased in a dose-dependent manner. Furthermore, the guanine-specific pattern of arrests produced when an exonuclease-free preparation of DNA polymerase beta was used corresponded exactly to the arrest patterns produced in vitro by the exonuclease-free enzyme Sequenase and ex vivo by Taq polymerase. These results suggest that replication arrest may be a common response of polymerases to DNA-chromium lesions and provide a plausible mechanism for the inhibition of DNA synthesis and S-phase cell-cycle delay that occurs in mammalian cells treated with genotoxic chromium compounds.

Topics: Ascorbic Acid; Cell Cycle; Cell Transformation, Neoplastic; Chromium; DNA Adducts; DNA Damage; DNA Polymerase beta; DNA Polymerase I; DNA Replication; DNA-Directed DNA Polymerase; DNA, Bacterial; Humans; Plasmids

The human hepatoma cells SMMC-7721 were treated with different concentrations of ascorbic acid (50-800 mumol/L) and FeSO4 (2.5-40 mumol/L) system to generate oxidative stress at various degrees. The oxidative stress induced by the system were mainly contributed to hydroxyl radical. All the various degrees of oxidative stress in this study are able to inhibit the proliferation of hepatoma cells. While low levels of oxidative stress may cause hepatoma cells lost some malignant features, such as aggregation of Con-A to the cell surface, alpha-fetoprotein, gamma-glutamyltransepeptidase and tyrosine-alpha-ketoglutarate transaminase, all of the 4 indices tended to cell differentiation, coloning efficiency potential decreased significantly, and apoptotic cells appeared. The numbers of apoptotic cells increased with the increasing of oxidative stress. The apoptotic cells exhibited non-adherent, smaller, chromatin condensed around the periphery of the nucleus in the shape of crescent, nuclear fragmentations but with intact cellular membrane, and DNA degraded to around 21.2 kbp fragment. All of the results showed that there is possibility to inhibit hepatoma cells growth, to promote differentiation and apoptosis, and therefore to initiate reverse transformation via strict regulation of oxidative stress.

Topics: Apoptosis; Ascorbic Acid; Carcinoma, Hepatocellular; Cell Division; Cell Transformation, Neoplastic; Free Radical Scavengers; Humans; Liver Neoplasms; Oxidative Stress; Tumor Cells, Cultured

The mechanisms of the tumor promoting activity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were studied using as in vitro model the enhancement ('promotion') of malignant transformation of C3H/M2 mouse fibroblasts induced by N-methyl-N'-nitro-N-nitrosoguanidine or 3-methylcholanthrene. In this assay, the promoting effect of TCDD was maximal at a very low concentration of 1.5 pM and was comparable to the effect of the reference tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA, 0.25 microg/ml). The role of reactive oxygen species in the promoting action was investigated: mannitol, a scavenger of hydroxyl radicals, or antioxidants, i.e. ascorbic acid plus alpha-tocopherol, abolished the in vitro promoting effects of TPA and TCDD. Furthermore, the involvement of protein kinase C (PKC) activation was studied: the protein kinase inhibitor H-7 markedly reduced the in vitro promoting activity of TPA but did not affect the promotion by TCDD. In accord with these results, TPA, but not TCDD, enhanced the PKC activity in C3H/M2 fibroblasts. Since the TPA-mediated activation of PKC was not affected by ascorbate plus alpha-tocopherol, it is concluded that the antioxidants interfere with tumor promotion at a step beyond PKC activation. Thus, the results suggest that the enhancement of malignant cell transformation by TPA and TCDD is dependent on a common mechanism, possibly induced by oxygen radicals, and, in addition, on further mechanisms that may involve agent-specific signalling pathways (e.g. PKC activation by TPA).

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Animals; Anticarcinogenic Agents; Antioxidants; Ascorbic Acid; Carcinogens; Cell Transformation, Neoplastic; Drug Interactions; Enzyme Inhibitors; Fibroblasts; Isoquinolines; Mannitol; Methylnitronitrosoguanidine; Mice; Mice, Inbred C3H; Piperazines; Polychlorinated Dibenzodioxins; Protein Kinase C; Reactive Oxygen Species; Tetradecanoylphorbol Acetate; Vitamin E

Vitamin C is an essential nutrient whose protective influence is carcinogenesis has been reported frequently, suggesting that vitamin C inhibits the formation of some carcinogens and decreases the incidence and delays of the neoplastic lesions. However, the mechanisms by which this occurs are unknown. In this study, the water soluble carcinogen 4-nitroquinoline-1-oxide (4NQO) has been used to induce a high yield of tumours in the oral cavity either singly or in combination with tobacco. Since the mucosa of rats is less susceptible to carcinogens than the hamster cheek pouch, the hamster cheek pouch has been used to study the influence of vitamin C on 4NQO-induced oral malignancy. The aim of this study was to determine whether topically applied vitamin C had an effect on the oral carcinogenesis induced by application of 4NQO. Similarly, an attempt was made to study the modulating effect of vitamin C on the histopathological and ultrastructural changes during the neoplastic process in the hamster. Vitamin C appeared to delay tumour induction and had other protective effects against neoplasia.

Topics: 4-Nitroquinoline-1-oxide; Animals; Ascorbic Acid; Carcinogens; Cell Transformation, Neoplastic; Cheek; Cricetinae; Mesocricetus; Microscopy, Electron; Mouth Neoplasms; Nicotiana; Plants, Toxic

The experiments reported here demonstrate that benzoyl peroxide (BP) can promote radiation induced transformation in vitro. BP is shown to be capable of generating free radicals, determined by the kinetics of hydroxylation as measured by fluorescence of coumarin-3-carboxylic acid. Although the mechanisms involved in the BP enhancement of radiation transformation are unknown, we hypothesize that lipid peroxidation produced by benzoyl radicals in the vicinity of membrane associated unsaturated lipids could contribute to the promotion of transformation in vitro.

Topics: Animals; Ascorbic Acid; Benzoyl Peroxide; Cell Line; Cell Transformation, Neoplastic; Dose-Response Relationship, Radiation; Free Radicals; Hydroxyl Radical; Hydroxylation; Iron; Kinetics; Mice; Radiation-Sensitizing Agents; Tetradecanoylphorbol Acetate; X-Rays

The effects of ascorbic acid on the neoplastic transformation induced by 60Co gamma-rays or 252Cf fission neutrons were studied. The transformation induced by each of the radiations was markedly suppressed when ascorbic acid was added daily to the culture medium during the first 8 days of the post-irradiation period. If ascorbic acid was added for a total of 8 days as mentioned above, but with 1 day interruption in the middle, the suppression of transformation was decreased. Ascorbic acid also suppressed the promotion of radiation transformation by 12-O-tetradecanoylphorbol-13-acetate (TPA) when both chemicals were added to the medium together. However, promotion of radiation transformation by TPA was observed if both the chemicals were added sequentially during the post-irradiation period, i.e. ascorbic acid for the first 7 days followed by TPA. These results suggest that continuous administration of ascorbic acid for a certain number of days is needed to suppress the radiation transformation, and also suggest that it acts on the promotion stage of transformation.

Topics: 3T3 Cells; Animals; Ascorbic Acid; Cell Transformation, Neoplastic; Cobalt Radioisotopes; Depression, Chemical; Gamma Rays; Mice; Neutrons

In primary avian tendon cells, Rous sarcoma virus can coexist or completely take over the cell. Infection, at high multiplicity or under conditions that promote high virus production (no ascorbate and high serum concentrations), results in almost complete oncogenic transformation of the culture. This is indicated in part by a radical change in morphology, growth at high cell density, and a dramatic drop in the production of procollagen from approximately 50% to approximately 3% of total protein synthesis. In contrast, infection at low multiplicity, infection with a replication defective virus, or the presence of ascorbate restrict the ability of the virus to transform the culture. Thus, there appears to be a balance between the normal and transformed states of the cell that can be shifted depending on the cellular environment and the level of infection. Ascorbate stabilizes the normal state by reducing virus production and promoting the synthesis of differentiated proteins.

Topics: Animals; Ascorbic Acid; Avian Sarcoma Viruses; Blood; Cell Differentiation; Cell Transformation, Neoplastic; Cells, Cultured; Culture Media; Sarcoma, Avian; Tendons; Virus Replication

Radioactivity measurements and autoradiographic studies of DNA, RNA and protein synthesis using [3H]-thymidine, [3H]-uridine and [3H]-leucine revealed that Vitamin C administration significantly (by 50%) decreases the DNA, RNA and protein synthesis in the neoplastic cells of basal cell carcinomas and squamous cell carcinomas in mice and rats. Basal cell carcinomas and squamous cell carcinomas were induced by a topical application of a chemical carcinogen, 3-methylcholanthrene. The inhibition of DNA, RNA and protein synthesis is accompanied by advanced ultrastructural and cell surface changes. Since the concentrations of Vitamin C were significantly higher in the plasma of Vitamin C-treated as compared to that of non Vitamin C-treated animals, these findings demonstrate that Vitamin C inhibits DNA, RNA and protein synthesis in neoplastic epithelial cells, and thus exerts its antineoplastic effect.

Topics: Animals; Ascorbic Acid; Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; DNA; Epithelium; Male; Methylcholanthrene; Mice; Protein Synthesis Inhibitors; Rats; Rats, Inbred Strains; RNA

Ascorbate irreversibly inhibited morphological transformation induced by methylcholanthrene in C3H/10T1/2 cells. To determine the mechanisms of this inhibition, we studied ascorbate uptake, redox potential, matrix proteins, and lipid composition of 10T1/2 cells. Ascorbate (16.8 nmol/dish) saturated cells and reduced the NADH-to-NAD+ ratio. Daily treatments with ascorbate, 28 nmol/dish, maintained intracellular ascorbate and reduced NADH by half. Matrix collagen and glycoproteins were stimulated by ascorbate, Iso-ascorbate, and dehydroascorbate in a dose-dependent manner. Both ascorbate and dehydroascorbate reduced total lipids with time; neutral lipids increased but were released into the media, phospholipids were modified, cholesterol-phospholipid ratios declined, and an inverse relationship between unsaturation index and cholesterol-phospholipids was apparent. Lipophilic bodies gradually accumulated. Our data suggest that inhibition of transformation by ascorbate, Iso-ascorbate, or dehydroascorbate may be associated with regulation of the redox potential, glycoproteins, and lipids in 10T1/2 cells.

Topics: Animals; Ascorbic Acid; Cell Line; Cell Transformation, Neoplastic; Dehydroascorbic Acid; Methylcholanthrene; NAD; Oxidation-Reduction

X-ray induced transformation of C3H10T1/2 cells was suppressed in a concentration-dependent manner by administration of ascorbic acid after irradiation (0.1-20 micrograms/ml for the first week) in the culture medium. The dose-response curve was shifted about 60% downward and was slightly steeper in the presence of ascorbic acid (5 micrograms/ml for the first week) than in its absence. The 1-week treatment procedure revealed that cells initiated by radiation remained susceptible to ascorbic acid until the time of morphological phenotype expression. The neoplastically transformed phenotype expressed after incubation for 8 weeks could no longer be suppressed by ascorbic acid even after culture transfer. Similarly, the neoplastically transformed phenotype suppressed for 8 weeks by ascorbic acid treatment was not subsequently expressed in the absence of ascorbic acid. On the basis of the oxygen-detoxifying nature of ascorbic acid, we postulated that expression of the neoplastically transformed phenotype is promoted by reactive oxygen species and peroxy radicals generated in cells during the whole assay period. The data may be useful as a guide for chemopreventive efforts against radiation carcinogenesis.

Topics: Animals; Ascorbic Acid; Cell Line; Cell Transformation, Neoplastic; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Mice

On the primary rat lung cell cultures, a study was made of the transforming action of sodium nitrite (NN) and amidopyrine combination with the control for formation of carcinogen--N-nitrosodimethylamine (NDMA) in growth medium. The manifestation of transformation was registered by appearance of morphological changes and multilayer growth foci. As criteria for evaluation after 48 hour treatments with NDMA, NN and AP, were used DNA-synthetizing activity of cells, mitotic index, frequency of mitoses pathology, monolayer density. The effects of transforming dose of NN alone and in combination with AP were the same. But malignization (tumor development in newborn rats in the points of cell suspension inoculation) took place only after administration of NN in combination with AP, when carcinogen was formed. Theophylline decreased the action of NN-AP combination.

Topics: Aminopyrine; Animals; Ascorbic Acid; Carcinogens; Cell Transformation, Neoplastic; Cells, Cultured; Lung; Mitosis; Nitrites; Nitroso Compounds; Rats; Sodium Nitrite; Theophylline

The relative amounts of iso-tRNAsGly and iso-tRNAsPro existing in chick embryo tendon are indicative of a specialization of the tRNA population for collagen synthesis. These amounts are not modified (i) in primary avian tendon (PAT) cells in culture for which the procollagen production varies from about 10% of total protein synthesis to 60% and (ii) in tendons from immature chicks, which show a 3-fold decrease of procollagen production with increasing age. The characteristic tRNA pattern was not maintained in cells which had lost the ability to make high levels of collagen as observed in the cases of: (i) PAT cells reaching confluency; (ii) virus-transformed PAT cells and (iii) tendon from adult chick. Our data are consistent with the idea that tendon tRNA specialization for collagen synthesis is a differentiation feature independent of the expression level of the collagenic function but related to its maintenance.

Topics: Aging; Animals; Ascorbic Acid; Avian Sarcoma Viruses; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Chickens; Kinetics; Procollagen; RNA, Transfer, Amino Acid-Specific; RNA, Transfer, Pro; Tendons

The effects of antioxidants on mammary gland carcinogenesis pretreated with 7,12-dimethylbenz[a]anthracene (DMBA) in female Sprague-Dawley rats were examined. The antioxidants used were butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), sodium L-ascorbate, alpha-tocopherol, ethoxyquin and p,p'-diaminodiphenylmethane (DDPM), which is an inhibitor of carcinogenesis in the liver, kidney and urinary bladder. Female Sprague-Dawley rats of 50 days old were treated with 2.5 mg/100 g body weight of DMBA, and from 1 week later were given diet supplemented with 1% BHA, 0.7% BHT, 5% sodium L-ascorbate, 1.5% alpha-tocopherol, 0.5% ethoxyquin or 0.1% DDPM for 33 weeks and then killed. The incidences of mammary tumors, carcinomas and fibroadenomas in DMBA-treated animals were reduced by diet containing BHA or ethoxyquin. Diet containing BHT or DDPM inhibited the induction of only fibroadenomas. The incidence of ear duct tumors in DMBA-treated animals was reduced by diet containing BHT, alpha-tocopherol or ethoxyquin.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Antioxidants; Ascorbic Acid; Bisacodyl; Butylated Hydroxyanisole; Butylated Hydroxytoluene; Cell Transformation, Neoplastic; Cresols; Diet; Ear Neoplasms; Ethoxyquin; Female; Mammary Neoplasms, Experimental; Rats; Rats, Inbred Strains; Vitamin E

Previous studies showing that prolyl hydroxylase is activated by addition of lactate to logarithmic cultures of L-929 and 3T6 mouse cell lines and that the enzyme normally increases in dense cultures which accumulate lactic acid, have led to the hypothesis that regulation is mediated through glycolytic metabolism. We present evidence that the extent of lactate production plays a minor role in controlling the density-dependent activation of prolyl hydroxylase in L-929 cells. The initial rate of glycolysis in stationary phase cells was lower than that in the logarithmic phase so that intracellular levels of glycolytic intermediates would not be related to activation of the enzyme. Nevertheless, because of increased numbers of cells in stationary phase, higher concentrations of lactate occur in the culture medium. Eliminating accumulation of lactate during the growth of L-929 cells by reducing the normal glucose concentration (5.5 mM) to low-glucose (0.5 mM) did not significantly affect the growth rate and still allowed a 10-fold activation of prolyl hydroxylase when cells reached stationary phase. Growth in normal glucose resulted in only an additional 2-fold increase in specific enzyme activity. These results imply that density-dependent activation of prolyl hydroxylase occurs mainly through a mechanism independent of lactate production. The elevated enzyme activity in dense cells was not decreased when lactate was removed and its production prevented by replacement of high-glucose medium with low-glucose medium for 24 hours. This result differed markedly from the dramatic decrease in enzyme activity observed within several hours after replating stationary phase cells in fresh medium, but at a lower cell density. The conclusion that there are at least two mechanisms regulating prolyl hydroxylase activity in L-929 cells was further supported by the observation that the maximal extent of enzyme activation by ascorbate or lactate in logarithmic phase L-929 cells was much less than the extent of density-dependent activation.

Topics: Animals; Ascorbic Acid; Cell Line; Cell Transformation, Neoplastic; Enzyme Activation; Glycolysis; Lactates; Lactic Acid; Mice; Procollagen-Proline Dioxygenase

C3H/10T 1/2 mouse embryo cells were transformed with 3-methylcholanthrene. Several type III morphologically transformed cell lines were selected with morphologies that either could or could not be reverted back to normal at passage 1 by daily addition of ascorbic acid (1 or 5 microgram/ml). Those transformed cell lines with morphologies that could be caused to revert to normal did not produce colonies in agarose or tumors in nude mice at early passages. Such transformed cell lines at later passages all formed colonies in agarose, but only 2 of 8 lines produced tumors at any passage tested. Subsequently, clones of transformed cells from each cell line have been isolated which are tumorigenic. In contrast, the transformed cell lines which were unresponsive to ascorbic acid at passage 1 were able to form colonies and to produce tumors in early passages. The reversion of the transformed morphology by ascorbic acid is apparently not caused by cytotoxicity since no cell kill was observed following exposure to ascorbate in any newly transformed cell lines at the concentrations used. Thus, the use of ascorbic acid allowed morphologically transformed cell lines to be isolated which appeared to be at different stages in the progression of an initiated cell from a morphologically transformed cell to a highly tumorigenic one. These studies also suggest that low concentrations of ascorbic acid in C3H/10T 1/2/CL8 cells can be effective in suppressing oncogenic progression only prior to a stage where an initiated cell achieves the capacity to grow in semisolid medium and to produce tumors in immunosuppressed animals. The importance of these cell lines for elucidating key changes required for the promotion and/or progression of cells to a tumorigenic phenotype is also presented.

Topics: Animals; Ascorbic Acid; Cell Division; Cell Line; Cell Survival; Cell Transformation, Neoplastic; Embryo, Mammalian; Methylcholanthrene; Mice; Mice, Inbred C3H; Mice, Nude; Mutation; Neoplasm Transplantation; Neoplasms; Neoplasms, Experimental; Phenotype

Topics: Adipose Tissue; Animals; Ascorbic Acid; Cell Division; Cell Transformation, Neoplastic; Collagen; Cyclic AMP; Extracellular Space; Methylcholanthrene; Mice; Phenotype

Ascorbic acid (50 micrograms/ml), added to the culture medium on a biweekly basis, suppresses the methylcholanthrene-induced transformation of C3H10T1/2 cells.

Topics: Animals; Ascorbic Acid; Cell Transformation, Neoplastic; Clone Cells; Methylcholanthrene

The BALB/c-3T3 cell neoplastic transformation system was modified to examine the tumor promoting activity of a set of substances. Following initiation of the target cells with 3-methylcholanthrene, treatment of the cultures with phorbol myristate acetate (0.01 microgram/ml; 1.5 X 10(-8) M) during the remainder of the 4-week assay interval resulted in a marked increase in both spontaneous and initiated Type III transformed foci. In contrast, a similar treatment with saccharin at 20, 100 or 500 microgram/ml (0.08, 0.4 or 2.1 X 10(-3) M) did not influence the occurrence of Type III transformed foci and did not result in a promoting response. Sodium ascorbate (2.53 X 10(-3) M) and L-tryptophan (2.45 X 10(-3) M) almost completely inhibited both spontaneous and initiated Type III transformed foci. Calcium pantothenate (2.10 X 10(-3) M) exhibited a marginal promoting effect. Under the conditions of this study in which the classical tumor promoter phorbol myristate acetate was highly active in promoting Type III transformed foci, saccharin was not active as either a direct transforming or promoting agent at doses up to 5 orders of magnitude higher.

Topics: Animals; Ascorbic Acid; Cell Transformation, Neoplastic; Cells, Cultured; Mice; Pantothenic Acid; Phorbols; Saccharin; Tetradecanoylphorbol Acetate; Tryptophan

Topics: Animals; Antineoplastic Agents; Ascorbic Acid; Cell Transformation, Neoplastic; Cells, Cultured; Drug Synergism; Drug Therapy, Combination; Glioma; Mice; Neoplasms, Experimental; Neuroblastoma; Vincristine; Vitamin E

Topics: Animals; Ascorbic Acid; Cell Transformation, Neoplastic; Cells, Cultured; Dose-Response Relationship, Drug; Embryo, Mammalian; Fibroblasts; Methylcholanthrene; Mice; Mice, Inbred C3H

Topics: Animals; Ascorbic Acid; Cannabis; Cell Transformation, Neoplastic; Cricetinae; Culture Media; Cysteine; DNA; Lung; Lysosomes; Nicotiana; Organ Culture Techniques; Plants, Toxic; Smoke

Collagen synthesis, hydroxylation of proline in collagen, and collagen secretion were studied in the contact-inhibited mouse fibroblast line, Balb 3T3; the Kirsten virus transformed line, Ki-3T3; and dibutyryl cAMP (dbcAMP)-treated Ki-3T3 cells, during the various phases of the growth cycle. Transformed cells in both logarithmic and stationary phase produced lower levels of collagen than the parent line but 85-90% of the theoretically possible hydroxyproline residues of the collagen were formed even when ascorbic acid was not added to the culture medium. Moreover, the transformed cells showed only about a 20% increase of collagen secretion upon addition of ascorbate. This was in contrast to the ascorbate requirement for maximal proline hydroxylation and the 2-3 fold stimulation of collagen secretion by ascorbate in the parent Balb 3T3 cells. Although dbcAMP treatment caused Ki-3T3 cells to assume a more normal morphology and increased the relative rate of collagen synthesis to levels similar to that of 3T3, such treatment did not restore an ascorbate requirement for proline hydroxylation or collagen secretion. The specific activity of the enzyme prolyl hydroxylase also was not affected by dbcAMP treatment although collagen synthesis was increased by such treatment. In addition, it was found that ascorbic acid was not effective in activating prolyl hydroxylase derived from Ki-3T3 or dbcAMP-treated Ki-3T3 cell cultures either in logarithmic phase or stationary phase. Ki-3T3 cultures did not accumulate ascorbic acid in cells or medium nor was ascorbic acid synthesized from the precursor 14C-glucuronate in cell homogenates. The results suggest that virally transformed Balb 3T3 cells acquire the capacity to synthesize a reducing cofactor for prolyl hydroxylase and that this function may be related to the increased glycolytic metabolism of these cells since neither cellular metabolism nor ascrobate-independent hydroxylation was altered by treatment with dbcAMP.

Topics: Ascorbic Acid; Bucladesine; Cell Line; Cell Transformation, Neoplastic; Collagen; Glucuronates; Hydroxylation; Procollagen-Proline Dioxygenase; Proline; Protein Biosynthesis

Topics: Animals; Ascorbic Acid; Carcinogens; Cell Transformation, Neoplastic; Mice; Nitrosamines