Journal Article•
Production of Large Amounts of Hydrogen Peroxide by Human Tumor Cells
TL;DR: Constitutive generation of large amounts of reactive oxygen intermediates, if it occurs in vivo, might contribute to the ability of some tumors to mutate, inhibit antiproteases, injure local tissues, and therefore promote tumor heterogeneity, invasion, and metastasis.
Abstract: Few nonphagocytic cells are known to generate reactive oxygen intermediates. Based on horseradish peroxidase-dependent, catalase-inhibitable oxidation of fluorescent scopoletin, seven human tumor cell lines constitutively elaborated H2O2 at rates (up to 0.5 nmol/10(4) cells/h) large enough that cumulative amounts at 4 h were comparable to the amount of H2O2 produced by phorbol ester-triggered neutrophils. Superoxide dismutase-inhibitable ferricytochrome c reduction was detectable at much lower rates. H2O2 production was inhibited by diphenyleneiodonium, a flavoprotein binder (concentration producing 50% inhibition, 0.3 microM), and diethyldithiocarbamate, a divalent cation chelator (concentration producing 50% inhibition, 3 microM), but not by cyanide or azide, inhibitors of electron transport, or by agents that inhibit xanthine oxidase, polyamine oxidase, or cytochrome P450. Cytochrome b559, present in human phagocytes and lymphocytes, was undetectable in these tumor cells by a sensitive spectrophotometric method. Mouse fibroblasts transfected with human tyrosinase complementary DNA made melanin, but not H2O2. Constitutive generation of large amounts of reactive oxygen intermediates, if it occurs in vivo, might contribute to the ability of some tumors to mutate, inhibit antiproteases, injure local tissues, and therefore promote tumor heterogeneity, invasion, and metastasis.
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TL;DR: There is growing evidence that aging involves, in addition, progressive changes in free radical-mediated regulatory processes that result in altered gene expression.
Abstract: At high concentrations, free radicals and radical-derived, nonradical reactive species are hazardous for living organisms and damage all major cellular constituents. At moderate concentrations, how...
9,131 citations
TL;DR: This review summarizes the current state of knowledge of the functions of NOX enzymes in physiology and pathology.
Abstract: For a long time, superoxide generation by an NADPH oxidase was considered as an oddity only found in professional phagocytes. Over the last years, six homologs of the cytochrome subunit of the phag...
5,873 citations
TL;DR: The spontaneous decay of DNA is likely to be a major factor in mutagenesis, carcinogenesis and ageing, and also sets limits for the recovery of DNA fragments from fossils.
Abstract: Although DNA is the carrier of genetic information, it has limited chemical stability. Hydrolysis, oxidation and nonenzymatic methylation of DNA occur at significant rates in vivo, and are counteracted by specific DNA repair processes. The spontaneous decay of DNA is likely to be a major factor in mutagenesis, carcinogenesis and ageing, and also sets limits for the recovery of DNA fragments from fossils.
5,209 citations
TL;DR: It is argued that modulating the unique redox regulatory mechanisms of cancer cells might be an effective strategy to eliminate these cells.
Abstract: Increased generation of reactive oxygen species (ROS) and an altered redox status have long been observed in cancer cells, and recent studies suggest that this biochemical property of cancer cells can be exploited for therapeutic benefits. Cancer cells in advanced stage tumours frequently exhibit multiple genetic alterations and high oxidative stress, suggesting that it might be possible to preferentially eliminate these cells by pharmacological ROS insults. However, the upregulation of antioxidant capacity in adaptation to intrinsic oxidative stress in cancer cells can confer drug resistance. Abrogation of such drug-resistant mechanisms by redox modulation could have significant therapeutic implications. We argue that modulating the unique redox regulatory mechanisms of cancer cells might be an effective strategy to eliminate these cells.
4,369 citations
TL;DR: It is argued that redox biology, rather than oxidative stress, underlies physiological and pathological conditions.
4,297 citations
Cites background from "Production of Large Amounts of Hydr..."
...Initial observations over two decades ago demonstrated that cancer cells generate higher levels of ROS than their non-transformed counterparts [35]....
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References
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TL;DR: O(2) (-) is made by leukocytes under circumstances which suggest that it may be involved in bacterial killing, and is identified as the agent responsible for the leukocyte-mediated reduction of cytochrome c.
Abstract: As a highly reactive substance produced in biological systems by the one-electron reduction of oxygen, superoxide (O(2) (-)) seemed a likely candidate as a bactericidal agent in leukocytes. The reduction of cytochrome c, a process in which O(2) (-) may serve as an electron donor, was found to occur when the cytochrome was incubated with leukocytes. O(2) (-) was identified as the agent responsible for the leukocyte-mediated reduction of cytochrome c by the demonstration that the reaction was abolished by superoxide dismutase, an enzyme that destroys O(2) (-), but not by boiled dismutase, albumin, or catalase. Leukocyte O(2) (-) production doubled in the presence of latex particles. The average rate of formation of O(2) (-) in the presence of these particles was 1.03 nmol/10(7) cells per 15 min. This rate, however, is only a lower limit of the true rate of O(2) (-) production, since any O(2) (-) which reacted with constituents other than cytochrome c would have gone undetected. Thus. O(2) (-) is made by leukocytes under circumstances which suggest that it may be involved in bacterial killing.
2,887 citations
Additional excerpts
...Appropriately stimulated PMNs3 (1-3) and monocytes (4-8) produce large amounts of ROI (up to 1.5 nmol of hydrogen peroxide/10" cells/h) in a respiratory burst pattern....
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...Appropriately stimulated PMNs3 (1-3) and monocytes (4-8)...
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TL;DR: This chapter describes two types of plasminogen activators—namely, the urokinase-type plasMinogen activator (u-PA) and the tissue- type plasmineg activator(t-PA), which are essentially different gene products.
Abstract: Publisher Summary This chapter discusses the role of plasminogen activators in various biological processes. In specific, it describes two types of plasminogen activators—namely, the urokinase-type plasminogen activator (u-PA) and the tissue-type plasminogen activator (t-PA), which are essentially different gene products. The amino acid sequences of these activators and nucleotide sequences of the corresponding cDNAs have largely been determined, and the cDNAs have been cloned using recombinant techniques. A variety of enzymatic as well as immunological assay and detection methods have also been developed that allows a precise quantification of the activators, a distinction between u-PA and t-PA, determination of whether an activator is present in its active or zymogen form, analysis of the kinetics of different steps of the cascade reaction, and immunocytochemical identification of u-PA and t-PA in tissue sections. Much of the studies on plasminogen activators and cancer has been guided by the hypothesis that proteolysis of the components of extracellular matrix, initiated by the release of plasminogen activator from the cancer cells, plays a decisive role for the degradation of normal tissue, and thereby for invasive growth and metastases.
2,545 citations
"Production of Large Amounts of Hydr..." refers background in this paper
...Plasmin ogen activator is produced by many tumors and is strongly implicated in tumorigenicity and metastasis (50-52)....
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TL;DR: Since Metschnikoff's discovery, hundreds of scientists studying dozens of species have reported thousands of studies on these cells, perhaps the most widely recognized of which are those of the eminent English scientists.
Abstract: (First of Two Parts) THE part played by phagocytes in defense against invading pathogens has been recognized since 1883. In that year, Metschnikoff, a Russian zoologist, reported that foreign particles injected into metazoans (in Metschnikoff's experiments, starfish larvae) were taken up by a population of "wandering mesodermal cells" that resided in interstitial tissues.1 He postulated a crucial role in host defense for these wandering cells, which he named "phagocytes." Since Metschnikoff's discovery, hundreds of scientists studying dozens of species have reported thousands of studies on these cells, perhaps the most widely recognized of which are those of the eminent English . . .
2,457 citations
Additional excerpts
...Appropriately stimulated PMNs3 (1-3) and monocytes (4-8) produce large amounts of ROI (up to 1.5 nmol of hydrogen peroxide/10" cells/h) in a respiratory burst pattern....
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...Appropriately stimulated PMNs3 (1-3) and monocytes (4-8)...
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01 Jan 1975
TL;DR: The careful attention to all the minute details provided by the dedicated tissue culturist seems to be the most important of all factors and cannot well be substituted for by special recipes or fancy equipment.
Abstract: There has been a feeling of frustration among many investigators trying to establish cell lines of human tumor cells. Numerous unsuccessful attempts, many of which have remained unpublished, have led to the conclusion that cell line establishment is controlled by the rare occurrence of a tumor with a built-in potential for long-term in vitro growth and by the application of intriguing culture techniques. The results given in the present chapter may, at least in part, contradict these conclusions. With techniques which in no particular way stand out as unusual, and by using medium and serum combinations, and in some cases added hormones, which are generally known to everyone involved in tissue culture, we have, during a rather short period of time, established a high number of new cell lines. These have originated from primary or metastatic human tumors, solid or effusions. In all fairness, many of our attempts to establish continuous lines have failed. However, the total number of attempts over successes is not truly representative of the frequency with which lines might be established. Many variations must be considered in this respect, including the choice of material, collection procedures, lapse of time between the clinical procedure and preparation for tissue culture, technical competence of assistants, and incidental factors known to everyone working in tissue culture. One thing, however, stands out as a conclusion: the careful attention to all the minute details provided by the dedicated tissue culturist seems to be the most important of all factors and cannot well be substituted for by special recipes or fancy equipment.
1,040 citations
"Production of Large Amounts of Hydr..." refers methods in this paper
...(26, 33) carcinoma cell lines were obtained from the late Dr....
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...The human colon carcinoma cell lines HT 29 (26) and HCT 15 (27) and the human pancreatic carcinoma cell line AsPC 1 (28, 29) were obtained from Dr....
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TL;DR: The results suggest that PMN adherent to intra- or extravascular surfaces may undergo a massive, prolonged respiratory burst at the command of macrophages and lymphocytes reacting to microbial products and antigens.
Abstract: Recombinant tumor necrosis factor alpha (rTNF alpha) and beta (rTNF beta) did not trigger H2O2 release from PMN in suspension. However, when PMN were plated on polystyrene surfaces coated with serum, fibronectin, vitronectin, laminin, or human umbilical vein endothelial cells (HUVEC), rTNFs induced a massive, prolonged secretory response, similar to that elicited by phorbol myristate acetate (PMA) or bacteria. On serum-coated plates, the maximum sustained rate of H2O2 release in response to rTNF alpha was 2.6 +/- 0.2 nmol/min per 10(6) PMN, the same as that with PMA; release continued for 73 +/- 4 min. On laminin-coated surfaces or HUVEC, release of H2O2 in response to rTNFs was slower, but lasted approximately 3.5 h, reaching the same total (greater than 100 nmol/10(6) PMN). Not only was this response far longer and larger than for other soluble stimuli of the respiratory burst studied with PMN in suspension, but the concentration necessary to elicit a half-maximal response (EC50) for rTNF alpha was orders of magnitude lower (55 pM). Responses were similar with FMLP, but ranged from zero to small with recombinant IFN alpha, recombinant IFN beta, recombinant IFN gamma, platelet-derived growth factor, recombinant IL-1 beta, or bacterial lipopolysaccharide. Adherent monocytes did not secrete H2O2 in response to rTNFs. H2O2 secretion by adherent PMN was first detectable 15-90 min after addition of rTNFs or FMLP. This lag period was unaffected by prior exposure of PMN to rTNF alpha in suspension, by allowing PMN to adhere before adding rTNF alpha, or by incubating adherent PMN in medium conditioned by rTNF alpha-treated PMN. Cytochalasins abolished H2O2 secretion in response to rTNFs, but not FMLP, if added during, but not after, the lag period. Thus, H2O2 secretion from rTNF alpha-treated PMN appears to be a direct but delayed response that requires assembly of microfilaments during exposure to the cytokine. These results suggest that PMN adherent to intra- or extravascular surfaces may undergo a massive, prolonged respiratory burst at the command of macrophages and lymphocytes reacting to microbial products and antigens.
809 citations
Additional excerpts
...5-10 MM),nor rTNF-a (10-300 ng/ml), agents capable of triggering a respiratory burst in PMNs in this system (35), could augment the observed H2O2 production rates in tumor cells as tested in SK-Mel 28, SK-Mel 30, SK-Mel 131, Lan 1, and HCT 15 cell...
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