Cancer incidence among nuclear workers in Russia based on data from the Institute of Physics and Power Engineering: a preliminary analysis.

One group that has the potential to be exposed to radiation is workers in the nuclear industry. Results of a systematic medical follow-up and dosimetric monitoring of these workers can form the basis for a study of the relationship between cancer incidence and radiation dose. As part of such efforts in Russia, a major institution of the nuclear industry with an established medical care unit, archiving capabilities, and dosimetry department was selected: the Institute of Physics and Power Engineering (IPPE) in Obninsk. In the study, a comparative analysis of cancer incidence rates for the IPPE workers and for the general population of Russia in 1991-1997 was carried out. The subjects were the IPPE workers hired before 1981. This restriction was imposed to reduce the uncertainty associated with the possible latent period in the development of solid cancers. Thus the possibility of including persons who already had the disease at the time when they were hired was minimized. The analysis is based on information about 158 cancer cases, including 24 cancers in persons under individual dosimetric monitoring. A statistically significant excess in cancer incidence was found among the IPPE workers compared with a comparison population (the general population of Russia) for some types of cancers. The SIR values for all cancers (ICD-9: 140-208) is 0.93 (95% CI 0.76, 1.12) for males and 1.42 (95% CI 1.06, 1.87) for females. A statistically significant excess for all cancers was also observed for residents of Obninsk compared to the control comparison population. The corresponding SIR value was 1.20 (95% CI 1.12, 1.28) for males and 1.58 (95% CI 1.49, 1.69) for females. An important reason for the observed excess in cancer incidence compared to the control population may be the higher level of health care in the so-called nuclear cities of Russia which may have resulted in increased diagnosis and registration of cancers. A statistically significant dependence of the cancer incidence on the dose of ionizing radiation was not established. The excess relative risk per gray for all types of cancer was 0.91 (95% CI -2.75, 4.61) for males and 0.40 (95% CI -6.94, 7.83) for females. These estimates should be considered to be preliminary, as the number of cases considered in the analysis of the dose response is small (17 males and 7 females).

Oncogenic Ras-induced proliferation requires autocrine fibroblast growth factor 2 signaling in skeletal muscle cells.

Constitutively activated Ras proteins are associated with a large number of human cancers, including those originating from skeletal muscle tissue. In this study, we show that ectopic expression of oncogenic Ras stimulates proliferation of the MM14 skeletal muscle satellite cell line in the absence of exogenously added fibroblast growth factors (FGFs). MM14 cells express FGF-1, -2, -6, and -7 and produce FGF protein, yet they are dependent on exogenously supplied FGFs to both maintain proliferation and repress terminal differentiation. Thus, the FGFs produced by these cells are either inaccessible or inactive, since the endogenous FGFs elicit no detectable biological response. Oncogenic Ras-induced proliferation is abolished by addition of an anti-FGF-2 blocking antibody, suramin, or treatment with either sodium chlorate or heparitinase, demonstrating an autocrine requirement for FGF-2. Oncogenic Ras does not appear to alter cellular export rates of FGF-2, which does not possess an NH(2)-terminal or internal signal peptide. However, oncogenic Ras does appear to be involved in releasing or activating inactive, extracellularly sequestered FGF-2. Surprisingly, inhibiting the autocrine FGF-2 required for proliferation has no effect on oncogenic Ras-mediated repression of muscle-specific gene expression. We conclude that oncogenic Ras-induced proliferation of skeletal muscle cells is mediated via a unique and novel mechanism that is distinct from Ras-induced repression of terminal differentiation and involves activation of extracellularly localized, inactive FGF-2.

ERK1/2 is required for myoblast proliferation but is dispensable for muscle gene expression and cell fusion.

Skeletal muscle satellite cells, which are found between the muscle fiber and the basal lamina, remain quiescent and undifferentiated unless stimulated to remodel skeletal muscle or repair injured skeletal muscle tissue. Quiescent satellite cells express c-met and fibroblast growth factor receptors (FGFR) 1 and 4, suggesting these receptors are involved in maintaining the undifferentiated quiescent state or involved in satellite cell activation. Although the signaling pathways involved are poorly understood, the mitogen activated protein kinase (MAPK) cascade has been implicated in the regulation of skeletal muscle growth and differentiation by FGFs. In this study, we investigated if activation of the Raf-MKK1/2-ERK1/2 signaling cascade plays a role in FGF-dependent repression of differentiation and proliferation of MM14 cells, a skeletal muscle satellite cell line. Inactivation ofthe Raf-MKK1/2-ERK1/2 pathway in myoblasts through the overexpression of dominant negative mutants of Raf-1 blocks ERK1/2 activity and prevents myoblast proliferation. Additionally, inhibition of MKK1/2 by treatment with pharmacological inhibitors also blocks FGF-mediated stimulation of ERK1/2 and blocks the G1 to S phase transition of myoblasts. Unexpectedly, we found that inactivation of the Raf-ERK pathway does not activate a muscle reporter, nor does inactivation of this pathway promote myogenic differentiation. We conclude that FGF-stimulated ERK1/2 signaling is required during the G1 phase of the cell cycle for commitment of myoblasts to DNA synthesis but is not required for mitosis once cells have entered the S-phase. Moreover, ERK1/2 signaling is not required either to repress differentiation, to promote skeletal muscle gene expression, or to promote myoblast fusion.

Regulation of myogenesis by fibroblast growth factors requires beta-gamma subunits of pertussis toxin-sensitive G proteins.

Terminal differentiation of skeletal muscle cells in culture is inhibited by a number of different growth factors whose subsequent intracellular signaling events are poorly understood. In this study, we have investigated the role of heterotrimeric G proteins in mediating fibroblast growth factor (FGF)-dependent signals that regulate myogenic differentiation. Pertussis toxin, which ADP-ribosylates and inactivates susceptible G proteins, promotes terminal differentiation in the presence of FGF-2, suggesting that Galpha or Gbeta gamma subunits or both are involved in transducing the FGF-dependent signal(s) that inhibits myogenesis. We found that Gbetagamma subunits are likely to be involved since the expression of the C terminus of beta-adrenergic receptor kinase 1, a Gbetagamma subunit-sequestering agent, promotes differentiation in the presence of FGF-2, and expression of the free Gbeta gamma dimer can replace FGF-2, rescuing cells from pertussis toxin-induced differentiation. Addition of pertussis toxin also blocked FGF-2-mediated activation of mitogen-activated protein kinases (MAPKs). Ectopic expression of dominant active mutants in the Ras/MAPK pathway rescued cells from pertussis toxin-induced terminal differentiation, suggesting that the Gbeta gamma subunits act upstream of the Ras/MAPK pathway. It is unlikely that the pertussis toxin-sensitive pathway is activated by other, as yet unidentified FGF receptors since PDGF (platelet-derived growth factor)-stimulated MM14 cells expressing a chimeric receptor containing the FGF receptor-1 intracellular domain and the PDGF receptor extracellular domain were sensitive to pertussis toxin. Our data suggest that FGF-mediated signals involved in repression of myogenic differentiation are transduced by a pertussis toxin-sensitive G-protein-coupled mechanism. This signaling pathway requires the action of Gbeta gamma subunits and activation of MAPKs to repress skeletal muscle differentiation.

The comparison of DNA synthesis inducibility by high potassium treatment in various nonproliferating cells.

Previous studies have demonstrated that treatment of fibroblasts from confluent, density-inhibited cultures with 50 mM KCl solution led to the onset of DNA replication. In the present study we show that such treatment can induce aphidicolin-sensitive DNA replication in differentiated nondividing cells: dorsal root ganglia neurons and in vitro differentiated myotubes formed by L6 myoblasts. Murine peritoneal macrophages and macrophages/granulocytes derived from human promyelocytic leukemia cells HL60 are refractory to high potassium treatment. These results confirm the nonuniformity of the nonproliferative state of various differentiated cells.

The short-term increase in extracellular potassium concentration causes DNA replication onset in density-inhibited cells.

The high potassium concentration effect on the human diploid fibroblasts (HDF) and 3T3 cells was investigated. The incubation of confluent cultures of HDF or 3T3 Swiss cells in the medium with 50 mM K+ for 35 min induced, 12 h later, the onset of DNA replication in a significant proportion of culture cell population. The same treatment had no effect upon the sparce cell cultures. No stimulation of DNA replication was observed in the absence of serum in culture medium.

Differential regulation of DNA synthesis in heterokaryons between chicken erythrocytes and culture cells with various proliferative potentials.

The regulation of DNA synthesis in heterokaryons between chicken erythrocytes and culture cells of various proliferative potential was studied. The following regularities were found: 1) Both immortalized and non-immortalized cells can efficiently reactivate DNA synthesis in erythrocyte nuclei. 2) Erythrocytes drastically inhibit the entry of non-malignant culture cell nuclei into the S-period, not acting upon DNA synthesis. 3) The inhibitory action is characteristic of erythrocytes from different stages of chicken ontogenesis (from 5-day-old embryos to the adult bird). 4) Malignant cells are completely refractory to the inhibitory action of erythrocytes. The ability of erythrocytes to inhibit the onset of replication in heterokaryons may be connected with the mechanisms of maintaining these terminally differentiated cells in a non-proliferating state.