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It is generally accepted that chronic inflammatory disease, either local or generalized, is associated with higher incidence of cancer. Since inflammation is often accompanied by oxidative stress the latter was indicated as the foundation for progressive mutations leading to tumor development (proliferation, invasion, metastasis). Even though, it is very hard to demonstrate by in vitro studies the causal relationship between oxidative stress and cell transformations. From our studies it is clear that cells are more likely to stop divisions and they commit suicide by apoptosis. During last decade, a novel view on the origin of cancer emerged. The so called cancer stem cells (CSC) were found that form the side-population of stem cells (SC) and they are believed to initiate cancer. Are the SC ancestors for CSC? Do SC transform into CSC? These and other questions remain unanswered. We hypothesize that SC might undergo transformation into CSC during prolonged oxidative stress. We claim that several changes in cell biochemistry has to occur to start the molecular modifications leading to neoplasma. These include either hypoxia-promoted apoptosis signal inducing kinase 1 (ASK-1), hypoxia inducing factor 1 alpha (HIF-la) and glycolysis, or normoxia-promoted activating protein-1 (AP-1) or hyperoxia-induced nuclear factor kappa B (NF-kB). Next, harsh microenviron- ment and heterogenous extracellular matrix (ECM) induced by oxidative stress accelerate the selection of clones of cells resistant to apoptogenic signal. HIF-la, protein crucial for transcriptional activation of protooncogene met leads to the overexpression of c-Met receptor that in turn sensitizes cells to hepatocyte growth factor/scatter factor (HGF/SF) mitogen. Finally, both impaired function of mitochondria and hypoxia elevate fibrin protein level and amplify hemostasis as disseminated in- tracapillary coagulation (DIC). In any case, it is very interesting and remains to be answered whether imbalance in prooxidant-antioxidant homeostasis has causal relationship with transformation of SC to CSC.
The article describes the principles of the insulin signaling pathway and the latest research results indicating the new role of insulin in modulating mitochondrial activity during muscle development. Myogenesis is considered to be an extensive energy-demanding process where mitochondria are the main source of ATP. Moreover, a number of reports emphasize that insulin is the most likely factor regulating prenatal muscle growth. Despite the fact that research into the affect of insulin onto myogenesis is incomplete, the role of mitochondria in muscle formation is thought to be essential in order to comprehend both whole-body growth and development. Insulin stimulates the expression of mitochondrial proteins in muscle cells whereas the activity of some targets of the insulin signaling pathway depends on ATP supply (e. g. mTOR). Similarly, alterations in available energy supply, resulting from the impaired function of mitochondria affect the cells sensitivity to insulin as well as leading to myopaties in the developing muscle tissue.
We previously demonstrated that both mitogenicity and anabolism in rat L6 myoblasts even in the presence of insulin could be markedly reduced by milimolar concentrations of H202, SIN-1 (3-mor- pholinosydnonimine) and SNP (sodium nitroprusside) - donors of peroxynitrite (ONOO ) and nitric oxide (NO), respectively (Orzechowski and Grzelkowska 2000). In this paper we described the results of the experiment, which was carried out in order to confirm the assumption that muscle cell death occurred after treatment with high (milimolar) doses of ROS/RNS. Hydrogen peroxide (H202) or SIN-1, or SNP, all induced cell death in rat L6 myoblasts when given at 1 mM. Symptoms of cell shrinking with nuclear chromatin condensation collapse of chromatin into patches along nuclear membrane and formation of apoptotic bodies occurred within 24 hours of the study. Apoptosis was evaluated in situ on the basis of apoptotic index. The process of oligonucleosomal fragmentation of nuclear DNA was demonstrated whenever apoptotic cell death was observed. The verification of cell viability was monitored and the above-mentioned index confirmed the lack of cell respiration in dead myoblasts, especially those treated with SNP. On average 92%, 80%, 77% and 65% of cells were found apoptotic after 1 mM H202, 1 mM SIN-1 or 0.5 mM SNP and 1 mM SNP, respectively. However, necrotic cell death significantly contributed to overall cell death in cultures treated with NO donor and amounted to 6% and 21% for 0.5 mM SNP and 1 mM SNP, respectively. Taken together, these results indicate massive apoptosis induced by H202 and SIN-1, but, with regard to apoptotic action of NO, cell respiration was additionally attenuated and associated with necrotic cell death.
This study investigates the effect of superoxide anion radical (02); hydrogen peroxide (H202), nitric oxide (NO) and peroxynitrite (ONOO), which often accompany inflamed, endotoxic or exercised muscle on insulin action in DTsatellite cells. In order to induce quiescence, rat L6 myoblasts were subjected to transition from G2/M to Gl phase by the application of serum-reduced medium. Insulin stimulating effect on cell mitogenicity and anabolism was dose-dependent and hormetic. Application of H202 alone enhanced protein synthesis with dose-dependency but had no effect on mitogenicity. While insulin and H202 were used together, (i.e. at low H202 dose) insulin action was not affected regardless of the combination used, except the loss of dose- dependency on protein synthesis, but for 100 μM of H202 insulin action ceased abruptly and totally. Since there were no additive effects of both factors, we conclude that H202 may contribute to the insulin-induced anabolic reaction, however, below 100 The application of 02- donor stimulated protein synthesis and slightly inhibited [cell proliferation] though dose-response pattern was not observed suggesting apparent limitations to 02- diffusion into the cell. Moreover 02- inhibited both insulin-enhanced mitogenicity and protein synthesis by abrogating dose-response fashion of insulin action. The introduction of NO and ONOO- donors alone to control systems inhibited cell proliferation in a dose-dependent manner having no effect on protein synthesis (except the low doses of SIN-1). Insulin-stimulated syntheses of both DNA and protein were inhibited in a dose- dependent manner by SIN-1 (NO and 02' donor). In the presence of SNP (NO donor) mitogenic effect of insulin was abolished whereas protein synthesis was diminished only by the highest SNP concentration used (0.5 mM). Taken together, these results have shown that hydrogen peroxide (H202), nitric oxide (NO) and peroxynitrite (ONOO ) provide a good explanation for developing resistance to growth promoting activity of insulin in satellite cells under conditions of oxidant stress.
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