8-hydroxyguanine and Colonic-Neoplasms

Altered oncogene expression in cancer cells causes loss of redox homeostasis resulting in oxidative DNA damage, e.g. 8-oxoguanine (8-oxoG), repaired by base excision repair (BER). PARP1 coordinates BER and relies on the upstream 8-oxoguanine-DNA glycosylase (OGG1) to recognise and excise 8-oxoG. Here we hypothesize that OGG1 may represent an attractive target to exploit reactive oxygen species (ROS) elevation in cancer. Although OGG1 depletion is well tolerated in non-transformed cells, we report here that OGG1 depletion obstructs A3 T-cell lymphoblastic acute leukemia growth in vitro and in vivo, validating OGG1 as a potential anti-cancer target. In line with this hypothesis, we show that OGG1 inhibitors (OGG1i) target a wide range of cancer cells, with a favourable therapeutic index compared to non-transformed cells. Mechanistically, OGG1i and shRNA depletion cause S-phase DNA damage, replication stress and proliferation arrest or cell death, representing a novel mechanistic approach to target cancer. This study adds OGG1 to the list of BER factors, e.g. PARP1, as potential targets for cancer treatment.

Topics: Animals; Antineoplastic Agents; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; DNA Damage; DNA Glycosylases; DNA Repair; DNA Replication; DNA, Neoplasm; Enzyme Inhibitors; Gene Expression Regulation, Neoplastic; Guanine; HCT116 Cells; Humans; Mice; Mice, Nude; Molecular Targeted Therapy; Oxidative Stress; Poly (ADP-Ribose) Polymerase-1; Reactive Oxygen Species; RNA, Small Interfering; Signal Transduction; Survival Analysis; Tumor Burden; Xenograft Model Antitumor Assays

Topics: Acetylcysteine; Animals; Colon; Colonic Neoplasms; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 2; DNA Damage; DNA Glycosylases; Glucose; Guanine; HCT116 Cells; HT29 Cells; Humans; Mice; Oxidative Stress

Oxidative DNA damage may be a risk factor for development of various pathologies, including malignancy. We studied inflammation triggered modulation of repair activity in the intestines of three weeks old rats injected i.p. with E.coli or S. typhimurium lipopolysaccharides (LPS) at doses of 1, 5 or 10 mg/kg. Subsequent formation in these animals of colonic preneoplastic lesions, aberrant crypt foci (ACF) was also investigated. Five days after LPS administration no differences were observed in repair rate of 1,N(6)-ethenoadenine (εA), 3,N(4)-ethenocytosine (εC) and 8-oxoguanine (8-oxoG) in intestines of these rats, as measured by the nicking assay. However a significant increase in all three repair activities was found within one and two months after S. typhimurium LPS treatment. E. coli LPS significantly increased only the 8-oxoG repair. S. typhimurium LPS stimulated mRNA transcription of pro-inflammatory proteins, lipooxygenase-12 and cyclooxygenase-2, as well as some DNA repair enzymes like AP-endonuclease (Ape1) and εC-glycosylase (Tdg). mRNA level of DNA glycosylases excising εA (MPG) and 8-oxoG (OGG1) was also increased by LPS treatment, but only at the highest dose. Transcription of all enzymes increased for up to 30 days after LPS, and subsequently decreased to the level observed before treatment, with the exception of APE1, which remained elevated even two months after LPS administration. Thus, the repair efficiency of εA, εC and 8-oxoG depends on the availability of APE1, which increases OGG1 and TDG turnover on damaged DNA, and presumably stimulates MPG. One and two months after administration of E. coli or S. typhimurium LPS, the number of aberrant crypt foci in rat colons increased in a dose and time dependent manner. Thus, inflammation stimulates the repair capacity for εA, εC and 8-oxoG, but simultaneously triggers the appearance of preneoplastic changes in the colons. This may be due to increased oxidative stress and imbalance in DNA repair.

Topics: Adenine; Animals; Animals, Newborn; Arachidonate 12-Lipoxygenase; Colon; Colonic Neoplasms; Cyclooxygenase 2; Cytosine; DNA Damage; DNA Repair; Escherichia coli; Guanine; Inflammation; Lipopolysaccharides; Oxidative Stress; Precancerous Conditions; Rats, Wistar; Salmonella typhimurium

The aim of this work was to answer the question whether the broad range of parameters which describe oxidative stress and oxidatively damaged DNA and repair are appropriate prognosis factors of colon cancer (CRC) patients survival? The following parameters were analyzed for 89 CRC patients: concentration of uric acid and vitamins A, E, C in plasma; levels of 8-oxodGuo (8-oxo-7,8-dihydro-2'-deoxyguanosine) in DNA of leukocyte and colon tissues; urinary excretion rates of 8-oxodGuo and 8-oxoGua (8-oxo-7,8-dihydroguanine); the activity and mRNA or protein level of repair enzymes OGG1, APE1, ANPG, TDG and PARP1. All DNA modifications and plasma antioxidants were analyzed using high performance liquid chromatography (HPLC) or HPLC/gas chromatography-mass spectrometry techniques. Expression of repair proteins was analyzed by QPCR, Western or immunohistochemistry methods. Longer survival coincided with low levels of 8-oxodGuo/8oxoGua in urine and 8-oxodGuo in DNA as well as with high concentration of uric acid plasma level. In contrast to expectations, longer survival coincided with lower mRNA level in normal colon tissue of the main 8-oxoGua DNA glycosylase, OGG1, but no association was found for PARP-1 expression. When analyzing simultaneously two parameters the discriminating power increased significantly. Combination of low level of urinary 8-oxoGua together with low level of 8-oxodGuo in leukocyte (both below median value) or high concentration of plasma uric acid (above median value) have the best prediction power. Since prediction value of these parameters seems to be comparable to conventional staging procedure, they could possibly be used as markers to predict clinical success in CRC treatment.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Case-Control Studies; Chromatography, High Pressure Liquid; Colonic Neoplasms; Deoxyguanosine; DNA Damage; DNA Repair Enzymes; Female; Follow-Up Studies; Gas Chromatography-Mass Spectrometry; Guanine; Humans; Male; Middle Aged; Neoplasm Staging; Oxidative Stress; Prognosis; Survival Rate; Uric Acid

Increased intestinal bile acids as a possible consequence of a high fat/meat, low fiber diet are believed to play an important role in the formation of colon cancer. Interactions of bile salts particularly secondary bile acids with different cell components including DNA may contribute to carcinogenesis. To further investigate DNA damage by bile salts, we assessed the effects of a bile salt mixture containing deoxycholate and chenodeoxycholate on base hydroxylation in Chelex-treated DNA from calf thymus as a model of human colonic mucosal DNA in the presence and absence of reactive oxygen metabolites (ROM).. Chelex-treated DNA from calf thymus (to remove residual iron impurities) was incubated with different bile salt concentrations (4 microM, 4.0 mM) (20.0% deoxycholate, 21.0% chenodeoxycholate) in the presence and absence of an OH generating system (25 microM FeCl3, 50 microM H2O2, 100 microM nitrilotriacetic acid) for 18 h (37 degrees C). After hydrolyzation, lyophilization and derivatization hydroxylated DNA bases were characterised and quantitated with gas chromatography-mass spectrometry (GS-MS) and SIM analysis. Two concentration ranges of bile salts were used, micromolar concentrations being present in plasma, millimolar in the gut lumen.. In the absence of ROM Chelex-treated DNA preparations contain only small amounts of hydroxylated base products. Bile salts at 4.0 mM significantly increased the amounts of 5-OH uracil and cis-thymine glycol. In the presence of ROM bile salts at 4.0 microM increased the production of 8-OH adenine and 8-OH guanine whereas bile salts at 4.0 mM inhibited ROM-induced base hydroxylation.. In the absence of ROM millimolar concentrations of a bile salt mixture with deoxycholate and chenodeoxycholate increase basal (spontaneous) DNA hydroxylation, whereas, they are without effects at micromolar concentrations. In the presence of ROM micromolar concentrations enhance oxidative DNA damage and millimolar concentrations were inhibitory. These results support the view that bile acids may cause oxidative DNA damage depending on their concentrations and the surrounding conditions both directly (enhancement of basal hydroxylation) and indirectly (enhancement of ROM-induced hydroxylation).

Topics: Adenine; Animals; Cattle; Chenodeoxycholic Acid; Colon; Colonic Neoplasms; Deoxycholic Acid; DNA; DNA Damage; Gas Chromatography-Mass Spectrometry; Gastrointestinal Agents; Guanine; Humans; Hydroxylation; Intestinal Mucosa; Models, Chemical; Reactive Oxygen Species; Salicylates; Thymus Gland

oh8Gua glycosylase repairs DNA by removing oh8Gua, a highly mutagenic oxidative DNA adduct. Recently, the gene for human oh8Gua glycosylase (hOGG1) was cloned and several mutational types have been reported. However, the implications of such mutations in human cancer have not been clearly demonstrated. To test the involvement of hOGG1 mutation in colon carcinogenesis, we analysed the genetic changes of hOGG1 and the activity of oh8Gua glycosylase in 15 paired normal and tumorous colon specimens. The activity of antioxidant enzymes (catalase and superoxide dismutase (SOD)) and extent of oxidative cellular damage (oh8Gua and malondialdehyde) were also assessed to compare the oxidative status of normal and tumour tissues. An Arg 154 to His mutation was detected in two tumour samples, but not in the corresponding normal tissues. A Ser 326 to Cys mutation (polymorphism) was found in both the normal and tumour tissues of 3 patients. However, neither the Arg 154 to His mutation nor the polymorphism at codon 326 significantly decreased the oh8Gua glycosylase activity. The mean activity of oh8Gua glycosylase was significantly higher in the tumours than in normal tissues (P=0.022). Antioxidant enzyme activities were decreased (catalase; P=0.004 and SOD; P=0.002), and the extent of oxidative damage correspondingly increased in the tumour tissues (oh8Gua; P=0.007 and malondialdehyde; P=0.046). Although the sample size was limited, these results suggest that the somatic mutation or the polymorphism of hOGG1 is less likely to be involved in colon carcinogenesis. Nevertheless, the greater oxidative DNA damage in the tumour tissues, as a possible result of impaired antioxidant activity, implies an important role for oxygen free-radicals in colon carcinogenesis.

Topics: Adult; Aged; Amino Acid Substitution; Colonic Neoplasms; DNA Adducts; DNA Damage; DNA-Formamidopyrimidine Glycosylase; Female; Guanine; Humans; Male; Middle Aged; Mutation; N-Glycosyl Hydrolases; Oxidative Stress