4-hydroxy-2-nonenal and Lung-Neoplasms

Manufacturers have developed prototype cigarettes yielding reduced levels of some tobacco smoke toxicants, when tested using laboratory machine smoking under standardised conditions. For the scientific assessment of modified risk tobacco products, tests that offer objective, reproducible data, which can be obtained in a much shorter time than the requirements of conventional epidemiology are needed. In this review, we consider whether biomarkers of biological effect related to oxidative stress can be used in this role. Based on published data, urinary 8-oxo-7,8-dihydro-2-deoxyguanosine, thymidine glycol, F2-isoprostanes, serum dehydroascorbic acid to ascorbic acid ratio and carotenoid concentrations show promise, while 4-hydroxynonenal requires further qualification.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Aldehydes; Ascorbic Acid; Biomarkers; Carotenoids; Dehydroascorbic Acid; Deoxyguanosine; F2-Isoprostanes; Humans; Lung Neoplasms; Oxidative Stress; Risk; Smoking; Thymidine; Tobacco Products; Tobacco Smoke Pollution

The aim of this study was to determine whether membrane lipid peroxidation in mammalian cells is enhanced by X-ray irradiation at the K-shell resonance absorption peak of phosphorus. A549 and wild-type p53-transfected H1299 (H1299/wtp53) cell lines derived from human lung carcinoma were irradiated with monoenergetic X-rays at 2.153 keV, the phosphorus K-shell resonance absorption peak, or those at 2.147 or 2.160 keV, which are off peaks. Immunofluorescence staining for 4-hydroxy-2-nonenal (HNE), a lipid peroxidation product, was used as marker for protein modification. In both cell lines, the HNE production was significantly enhanced after irradiation at 2.153 keV compared to sham-irradiation. The enhancement (E) was calculated as the ratio of the fluorescence intensity of irradiated cells to that of sham-irradiated cells. In both the cell lines, E2.153 was significantly larger than E2.147 and no significant difference between E2.147 and E2.160 was observed. The extra enhancement at 2.153 keV was possibly caused by energy transition within the phosphorus K-shell resonance absorption. Our results indicate that membrane lipid peroxidation in cells is enhanced by the Auger effect after irradiation at the K-shell resonance absorption peak of phosphorus rather than by the photoelectric effect of the constituent atoms in the membrane lipid at 2.147 keV.

Topics: Aldehydes; Cell Line, Tumor; Cell Membrane; Fluorescence; Humans; Lipid Peroxidation; Lung Neoplasms; Phosphorus; Radiation Dosage; X-Rays

The metabolic function of peroxisome proliferator-activated receptor gamma (PPARγ) in lung cancer remains unclear.. To determine the relationship of PPARγ on ALDH1A3-induced lipid peroxidation to inhibit lung cancer cell growth.. In silico analysis using microarray dataset was performed to screen the positive correlation between PPARγ and all ALDH isoforms. NUBIscan software and ChIP assay were used to identify the binding sites (BSs) of PPARγ on ALDH1A3 promoter. The expression of ALDH1A3 under thiazolidinedione (TZD) treatment was evaluated by QPCR and Western Blot in HBEC and H1993 cell lines. Upon treatment of TZD, colony formation assay was used to check cell growth inhibition and 4-hydroxy-2-nonenal (4HNE) production as lipid peroxidation marker was determined by Western Blot in PPARγ positive cell H1993 and PPARγ negative cell H1299.. Compared to other ALDH isoforms, ALDH1A3 showed the highest positive correlation to PPARγ expression. ALDH1A3 upregulated PPARγ expression while PPARγ activation suppressed ALDH1A3. Among 2 potential screened PPARγ response elements, BS 1 and 2 in the promoter of ALDH1A3 gene, PPARγ bound directly to BS2. Ligand activation of PPARγ suppressed mRNA and protein expression of ALDH1A3. Growth inhibition was observed in H1993 (PPARγ positive cell) treated with PPARγ activator and ALDH inhibitor compared to H1299 (PPARγ negative cell). PPARγ activation increased 4HNE which is known to be suppressed by ALDH1A3.. ALDH1A3 suppression could be one of PPARγ tumor suppressive function. This study provides a better understanding of the role of PPARγ in lung cancer.

Topics: Aldehyde Oxidoreductases; Aldehydes; Apoptosis; Binding Sites; Cell Line, Tumor; Cell Proliferation; Gene Expression Regulation, Neoplastic; Humans; Lipid Peroxidation; Lung Neoplasms; PPAR gamma; Protein Binding; Thiazolidinediones

The Aim of the study was to reveal if PET-CT analysis of primary and of secondary lung cancer could be related to the onset of lipid peroxidation in cancer and in surrounding non-malignant lung tissue.. Nineteen patients with primary lung cancer and seventeen patients with pulmonary metastasis were involved in the study. Their lungs were analyzed by PET-CT scanning before radical surgical removal of the cancer. Specific immunohistochemistry for the major bioactive marker of lipid peroxidation, 4-hydroxynonenal (HNE), was done for the malignant and surrounding non-malignant lung tissue using genuine monoclonal antibody specific for the HNE-histidine adducts.. Both the intensity of the PET-CT analysis and the HNE-immunohistochemistry were in correlation with the size of the tumors analyzed, while primary lung carcinomas were larger than the metastatic tumors. The intensity of the HNE-immunohistochemistry in the surrounding lung tissue was more pronounced in the metastatic than in the primary tumors, but it was negatively correlated with the cancer volume determined by PET-CT. The appearance of HNE was more pronounced in non-malignant surrounding tissue than in cancer or stromal cells, both in case of primary and metastatic tumors.. Both PET-CT and HNE-immunohistochemistry reflect the size of the malignant tissue. However, lipid peroxidation of non-malignant lung tissue in the vicinity of cancer is more pronounced in metastatic than in primary malignancies and might represent the mechanism of defense against cancer, as was recently revealed also in case of human liver cancer.

Topics: Adult; Aged; Aldehydes; Female; Humans; Immunohistochemistry; Lipid Peroxidation; Lung; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Positron Emission Tomography Computed Tomography

Lung cancer is the leading cause of cancer death worldwide and its clinical management remains challenge. Here, we repurposed antibiotic levofloxacin for lung cancer treatment. We show that levofloxacin is effectively against a panel of lung cancer cell lines via inhibiting proliferation and inducing apoptosis, regardless of cellular origin and genetic pattern, in in vitro cell culture system and in vivo xenograft lung tumor model. Mechanistically, levofloxacin inhibits activities of mitochondrial electron transport chain complex I and III, leading to inhibition of mitochondrial respiration and reduction of ATP production. In addition, levofloxacin significantly increases levels of ROS, mitochondrial superoxide and hydrogen peroxide in vitro and oxidative stress markers (HEL and 4-HNE) in vivo. Antioxidants, such as NAC and vitamin C, prevent the inhibitory effects of levofloxacin, confirming the induction of oxidative damage as the mechanism of its action in lung cancer cells. Our work demonstrates that levofloxacin is a useful addition to the treatment of lung cancer. Our work also suggests that targeting mitochondria may be an alternative therapeutic strategy for lung cancer treatment.

Topics: Aldehydes; Animals; Antibiotics, Antineoplastic; Apoptosis; Cell Line, Tumor; Cell Proliferation; DNA Damage; Dose-Response Relationship, Drug; Electron Transport Complex I; Electron Transport Complex III; Humans; Levofloxacin; Lung Neoplasms; Mice, SCID; Mitochondria; Oxidative Stress; Reactive Oxygen Species; Tumor Burden; Xenograft Model Antitumor Assays

The expression levels of human antioxidant genes (HAGs) and oxidative markers were investigated in light of lung adenocarcinoma aggressiveness and patient outcome.. We assayed in vitro the tumoral invasiveness and multidrug resistance in human lung adenocarcinoma (AdC) cell lines (EKVX and A549). Data were associated with several redox parameters and differential expression levels of HAG network. The clinicopathological significance of these findings was investigated using microarray analysis of tumor tissue and by immunohistochemistry in archival collection of biopsies.. An overall increased activity (expression) of selected HAG components in the most aggressive cell line (EKVX cells) was observed by bootstrap and gene set enrichment analysis (GSEA). In vitro validation of oxidative markers revealed that EKVX cells had high levels of oxidative stress markers. In AdC cohorts, GSEA of microarray datasets showed significantly high levels of HAG components in lung AdC samples in comparison with normal tissue, in advanced stage compared with early stage and in patients with poor outcome. Cox multivariate regression analysis in a cohort of early pathologic (p)-stage of AdC cases showed that patients with moderate levels of 4-hydroxynonenal, a specific and stable end product of lipid peroxidation, had a significantly less survival rate (hazard ratio of 8.87) (P < 0.05).. High levels of oxidative markers are related to tumor aggressiveness and can predict poor outcome of early-stage lung adenocarcinoma patients.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Adult; Aged; Aldehydes; Antioxidants; Cell Line, Tumor; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Lipid Peroxidation; Lung Neoplasms; Male; Microarray Analysis; Middle Aged; Multivariate Analysis; Neoplasm Invasiveness; Neoplasm Staging; Odds Ratio; Oxidation-Reduction; Predictive Value of Tests; Prognosis; Proportional Hazards Models; Retrospective Studies

Asbestos is a potent carcinogen associated with increased risks of malignant mesothelioma and lung cancer in humans. Although the mechanism of carcinogenesis remains elusive, the physicochemical characteristics of asbestos play a role in the progression of asbestos-induced diseases. Among these characteristics, a high capacity to adsorb and accommodate biomolecules on its abundant surface area has been linked to cellular and genetic toxicity. Several previous studies identified asbestos-interacting proteins. Here, with the use of matrix-assisted laser desorption ionization-time of flight mass spectrometry, we systematically identified proteins from various lysates that adsorbed to the surface of commercially used asbestos and classified them into the following groups: chromatin/nucleotide/RNA-binding proteins, ribosomal proteins, cytoprotective proteins, cytoskeleton-associated proteins, histones and hemoglobin. The surfaces of crocidolite and amosite, two iron-rich types of asbestos, caused more protein scissions and oxidative modifications than that of chrysotile by in situ-generated 4-hydroxy-2-nonenal. In contrast, we confirmed the intense hemolytic activity of chrysotile and found that hemoglobin attached to chrysotile, but not silica, can work as a catalyst to induce oxidative DNA damage. This process generates 8-hydroxy-2'-deoxyguanosine and thus corroborates the involvement of iron in the carcinogenicity of chrysotile. This evidence demonstrates that all three types of asbestos adsorb DNA and specific proteins, providing a niche for oxidative modification via catalytic iron. Therefore, considering the affinity of asbestos for histones/DNA and the internalization of asbestos into mesothelial cells, our results suggest a novel hypothetical mechanism causing genetic alterations during asbestos-induced carcinogenesis.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Aldehydes; Animals; Asbestos, Amosite; Asbestos, Crocidolite; Asbestos, Serpentine; Chromatin; Cytoskeleton; Deoxyguanosine; DNA; DNA Damage; Hemoglobins; Histones; Iron; Lung Neoplasms; Mesothelioma; Mice; Oxidation-Reduction; Proteins; Rats; Ribosomal Proteins; RNA-Binding Proteins; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Surface Properties

Multidrug-resistant proteins (MRPs) are members of the ATP-binding cassette superfamily that facilitate detoxification by transporting toxic compounds, including chemotherapeutic drugs, out of cells. Chemotherapy, radiation, and other xenobiotic stresses have been shown to increase levels of select MRPs, although the underlying mechanism remains largely unknown. Additionally, MRP3 is suspected of playing a role in the drug resistance of non-small-cell lung carcinoma (NSCLC). Analysis of the MRP3 promoter revealed the presence of multiple putative electrophile-responsive elements (EpREs), sequences that suggest possible regulation of this gene by Nrf2, the key transcription factor that binds to EpRE. The goal of this investigation was to determine whether MRP3 induction was dependent upon the transcription factor Nrf2. Keap1, a key regulator of Nrf2, sequesters Nrf2 in the cytoplasm, preventing entry into the nucleus. The electrophilic lipid peroxidation product 4-hydroxy-2-nonenal (HNE) has been shown to modify Keap1, allowing Nrf2 to enter the nucleus. We found that HNE up-regulated MRP3 mRNA and protein levels in cell lines with wild-type Keap1 (the human bronchial epithelial cell line HBE1 and the NSCLC cell line H358), but not in the Keap1-mutant NSCLC cell lines (A549 and H460). Cell lines with mutant Keap1 had constitutively higher MRP3 that was not increased by HNE treatment. In HBE1 cells, silencing of Nrf2 with siRNA inhibited induction of MRP3 by HNE. Finally, we found that silencing Nrf2 also increased the toxicity of cisplatin in H358 cells. The combined results therefore support the hypothesis that MRP3 induction by HNE involves Nrf2 activation.

Topics: Aldehydes; Base Sequence; Bronchi; Carcinoma, Non-Small-Cell Lung; Cisplatin; Epithelium; Gene Expression Regulation, Neoplastic; Gene Silencing; Humans; Lung Neoplasms; Molecular Sequence Data; Multidrug Resistance-Associated Proteins; NF-E2-Related Factor 2; Promoter Regions, Genetic; RNA, Messenger; RNA, Small Interfering; Tumor Cells, Cultured; Up-Regulation

RLIP76 functions as an ATP-dependent transporter of amphiphilic chemotherapeutic drugs such as doxorubicin (DOX, adriamycin), as well as of glutathione-conjugates of endogenous electrophilic toxins such as 4-hydroxynonenal (4HNE). RLIP76 couples transport and ATP-hydrolysis with a 1:1 stoichiometry, making the ATPase activity of RLIP76 an excellent surrogate for its transport activity. Present studies were performed to determine the relationship of the RLIP76 ATPase activity with DOX and 4HNE resistance in a panel of 13 native human lung cancer cell lines. RLIP76 was purified from each cell line and homogeneity demonstrated by SDS-PAGE and amino acid composition analysis. Anti-RLIP76 antibodies were shown by Ouchterlony double immunodiffusion tests to be non-cross-reactive with any other proteins including P-glycoprotein (Pgp) or multidrug resistance associated protein (MRP). These antibodies completely immunoprecipitated ATPase activity of purified RLIP76 fractions, further confirming homogeneity of purified RLIP76. RLIP76 ATPase purified from NSCLC cell lines was about 2-fold more active than that from SCLC in the absence of the stimulator dinitrophenyl S-glutathione (206+/-47, n=7 vs. 94+/-22, n=6, nmol/min/mg protein, respectively), or in its presence (340+/-60, n=7 vs. 186+/-32, n=6, nmol/min/mg; p<0.01). Partial tryptic digest revealed a 44 kDa internal fragment of RLIP76 beginning at Thr-294 in NSCLC cell lines. This fragment was absent from all SCLC, suggesting the possibility that the activity of RLIP76 in SCLC and NSCLC is differentially regulated through post-translational modifications. Taken together, these findings suggest that RLIP76 activity is a general determinant of 4HNE and DOX resistance, and that its activity contributes to the drug-resistant phenotype of NSCLC.

Topics: Adenosine Triphosphatases; Aldehydes; Amino Acid Sequence; Antineoplastic Agents; ATP-Binding Cassette Transporters; Biological Transport; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carrier Proteins; Cross Reactions; Doxorubicin; Drug Resistance, Neoplasm; Glutathione; GTPase-Activating Proteins; HL-60 Cells; Humans; Immunoglobulin G; K562 Cells; Lung Neoplasms; Molecular Sequence Data; Neoplasm Proteins; Protein Processing, Post-Translational; Trypsin; Tumor Cells, Cultured; U937 Cells

In A549 cell culture, significant variability was found in sensitivity to actinomycin D. Using limiting dilution, actinomycin D-susceptible (G4S) and -resistant (D3R) subclones were isolated. G4S cells were also susceptible to protein synthesis inhibitors, a redox cycling quinone, and an electrophile with concomitant activation of caspases 3 and 9. D3R cells were resistant to these agents without caspase activation. Antioxidant profiles revealed that D3R cells had significantly higher glutathione and glutathione reductase activity but markedly lower catalase, glutathione peroxidase, and aldehyde reductase activities than G4S cells. Thus A549 cells contain at least two distinct subpopulations with respect to predisposition to cell death and antioxidant profile. Because sensitivities to agents and the antioxidant profile were inconsistent, mechanisms independent of antioxidants, including the apparent inability to activate caspases in D3R cells, may play an important role. Regardless, the results suggest that antioxidant profiles of asymmetrical cell populations cannot predict sensitivity to oxidants and warn that the use of single subclones is advisable for mechanistic studies using A549 or other unstable cell lines.

Topics: Adenocarcinoma; Aldehydes; Anisomycin; Antioxidants; Apoptosis; Caspases; Cytotoxins; Dactinomycin; Genetic Heterogeneity; Glutathione; Growth Inhibitors; Humans; Hydrogen Peroxide; Lung Neoplasms; Male; Naphthoquinones; Oxidation-Reduction; Oxidative Stress; Protein Synthesis Inhibitors; Superoxide Dismutase; Tumor Cells, Cultured

In experiments using the renal carcinogen ferric nitrilotriacetate (Fe-NTA) in male ddY mice, primary pulmonary cancers were also induced in bronchiolar and alveolar tissues. 4-Hydroxy-2-nonenal (4-HNE) and 8-hydroxy-2'-deoxyguanosine (8-OHdG), products of oxidative processes, increased in bronchiolar and alveolar cells after administration of Fe-NTA. These substances disappeared after oral administration of propolis or artepillin C, as shown histochemically, and correlated with an anticancer prophylactic effect of propolis and artepillin C. From our investigation, lipid peroxidation seems to play an important role in pulmonary carcinogenesis. Malignant progression from adenoma of bronchiolar or alveolar origin to malignant tumors has been proposed to involve a stepwise transformation. In our study, adenomas developed into adenocarcinomas and large cell carcinomas after treatment with Fe-NTA. In contrast, after oral administration of propolis or artepillin C, adenomas did not progress to carcinomas. Instead of developing into large cell cancers, as induced by Fe-NTA in control mice, adenomas showed remarkable proliferation of macrophages and local anti-oxidant activity after treatment with either propolis or artepillin C. Propolis and artepillin C therefore appear to inhibit lipid peroxidation and the development of pulmonary cancers.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Aldehydes; Animals; Antineoplastic Agents; Deoxyguanosine; Ferric Compounds; Immunohistochemistry; Lipid Peroxidation; Lung Neoplasms; Male; Mice; Nitrilotriacetic Acid; Nuclear Proteins; Phenylpropionates; Proliferating Cell Nuclear Antigen; Propolis; Thyroid Nuclear Factor 1; Transcription Factors

4-Hydroxynonenal (4HNE) is the most prevalent toxic lipid peroxidation product formed during oxidative stress. It exerts its cytotoxicity mainly by the modification of intracellular proteins. The detection of 4HNE-modified proteins in several degenerative disorders suggests a role for 4HNE in the onset of these diseases. Efficient protection mechanisms are required to prevent the intracellular accumulation of 4HNE. The toxicity of 4HNE was tested with the small cell lung cancer cell lines GLC(4) and the multidrug-resistance-protein (MRP1)-overexpressing counterpart GLC(4)/Adr. In the presence of the MRP1 inhibitor MK571 or the GSH-depleting agent buthionine sulphoximine, both cell lines became more sensitive and showed decreased survival. Transport experiments were performed with the (3)H-labelled glutathione S-conjugate of 4HNE ([(3)H]GS-4HNE) with membrane vesicles from GLC(4)-derived cell lines with different expression levels of MRP1. [(3)H]GS-4HNE was taken up in an ATP-dependent manner and the transport rate was dependent on the amount of MRP1. The MRP1 inhibitor MK571 decreased [(3)H]GS-4HNE uptake. MRP1-specific [(3)H]GS-4HNE transport was demonstrated with membrane vesicles from High Five insect cells overexpressing recombinant MRP1. Kinetic experiments showed an apparent K(m) of 1.6+/-0.21 microM (mean+/-S.D.) for MRP1-mediated [(3)H]GS-4HNE transport. In conclusion, MRP1 has a role in the protection against 4HNE toxicity and GS-4HNE is a novel MRP1 substrate. MRP1, together with GSH, is hypothesized to have a role in the defence against oxidative stress.

Topics: Adenosine Triphosphate; Aldehydes; Animals; ATP Binding Cassette Transporter, Subfamily B, Member 1; ATP-Binding Cassette Transporters; Biological Transport; Buthionine Sulfoximine; Carcinoma, Small Cell; Cell Line; Cell Survival; Cysteine Proteinase Inhibitors; Dose-Response Relationship, Drug; Enzyme Inhibitors; Glutathione; Humans; Immunoblotting; Insecta; Kinetics; Leukotriene Antagonists; Lipid Peroxidation; Lung Neoplasms; Multidrug Resistance-Associated Proteins; Oxidative Stress; Propionates; Quinolines; Recombinant Proteins; Time Factors; Tumor Cells, Cultured

Trans-4-hydroxy-2-nonenal (HNE) is a product of lipid peroxidation. In the presence of t-butyl hydroperoxide the racemic HNE readily converts to its epoxide, 2,3-epoxy-4-hydroxynonanal (EH), as a pair of diastereomers. In this study, the potential roles of HNE and EH as tumor initiating agents were assessed. The mutagenicities of HNE and EH isomers in Salmonella strains TA100 and 104 were examined. In addition, the tumor initiating activities of HNE and EH were evaluated in bioassays involving either topical application in CD-1 mice or i.p. administration in newborn CD-1 mice. In the mutagenicity assays, EH isomers induced similar levels of revertants in both tester strains, although EG isomers were previously shown to react with bases in DNA with different specificity (Sodum, R.S. and Chung, F.-L., Cancer Res., 51, 137-143, 1991). The major isomer induced approximately 20,000 revertants/mumol in TA100 and 15,000 revertants/mumol in TA104, whereas, the minor isomer induced approximately 40,000 revertants/mumol in TA100 and 20,000 revertants/mumol in TA104. HNE was, however, not mutagenic under the assay conditions. In the tumor bioassays, EH was a weak tumorigen in CD-1 mice upon topical application followed by TPA promotion, yielding 0.55 tumors/mouse and 40% tumor incidence at a total dose of 128 mumol/mouse versus 0.02 tumors/mouse and 5% tumor incidence in the control group. Both HNE and EH induced liver tumors in male mice, but not in female mice. However, the incidences were not statistically significant. EH administered i.p. at a total dose of 200 nmol/mouse exacerbated the chronic spontaneous nephropathy in newborn CD-1 mice. Although the incidence of mild nephropathy was comparable in both EH-treated and control groups, the incidence of more severe lesions in mice treated with 200 nmol/mouse was 21%; while it was 0% in the control group. Furthermore, two mice at each dose level of EH showed a tubule profile with complex hyperplastic lining, suggestive of atypical hyperplasia. Again, HNE was not as active as EH in these bioassays. These results suggest a possible role of EH in tumorigenesis associated with lipid peroxidation.

Topics: Aldehydes; Animals; Animals, Newborn; Epoxy Compounds; Female; Kidney; Lipid Peroxidation; Liver Neoplasms, Experimental; Lung Neoplasms; Male; Mice; Mutagenicity Tests; Salmonella; Skin Neoplasms