Neoplastic transformation of human cells

The goal of this project was to gain a better understanding of the cellular mechanisms of cancer induction by ionizing radiation as a risk assessment for workers subjected to high LET irradiation such as that found in space. The following ions were used for irradiation: Iron, Argon, Neon, and Lanthanum. Two tests were performed: growth in low serum and growth in agar were used as indicators of cell transformation. The specific aims of this project were to: (1) compare the effectiveness of various ions on degree of transformation of a single dose of the same RBE; (2) determine if successive irradiations with the same ion (Ge 600 MeV/u) increases the degree of transformation; (3) test if clones with the greatest degree of transformation produce tumors in nude mice; and (4) construct a cell hybrid of a transformed and control (non-transformed) clone. The cells used for this work are human mammary epithelial cells with an extended lifespan and selected for growth in MEM + 10% serum

Increased cytotoxicity of oxidized flame soot

AbstractCombustion–generated particles released into the atmosphere undergo reactions with oxidants, which can change the particles’ physiochemical characteristics. In this work, we compare the physical and chemical properties and cellular response of particles fresh from a flame with those oxidized by ozone and nitrogen dioxide. The reaction with ozone and nitrogen dioxide does not significantly modify the physical characteristics of the particles (primary particle size, fractal dimension, and surface area). However, oxidation affects the chemical characteristics of the particles, creating more oxygen and nitrogen containing functional groups, and increases their hydrophilicity. In addition, oxidized soot generates more reactive oxygen species, as measured by the dithiothreitol (DTT) assay. Furthermore, oxidized soot is 1.5 – 2 times more toxic than soot that was not reacted with ozone, but the inflammatory response, measured by interleukin–8 (IL–8) secretion, is unchanged. These results imply that combustion–generated particles released into the atmosphere will have an increased toxicity on or after high ozone days

Molecular and cellular mechanisms in nervous system-specific carcinogenesis by N-ethyl-N-nitrosourea

A single pulse of N-ethyl-N-nitrosourea (ENU), applied to BDIX rats during the perinatal age, specifically results in a high incidence of neuroectodermal neoplasms in the central and peripheral nervous system (NS). The pronounced sensitivity of the developing NS suggests a dependence of the carcinogenic effect on the proliferative and/or differentiative state of the target cells at the time of the ENU pulse. The specificity of ENU for the NS cannot be due to tissue variations in the degree of carcinogen-cell interactions, since the reactive, electrophilic ethyl cation is produced by rapid, nonenzymatic decomposition of ENU indiscriminately in all tissues. Correspondingly, the initial molar fractions of ethylated purine bases are similar in the DNA of "high-risk" (perinatal brain) and "low-risk" tissues (e.g., liver; adult brain). However, while the respective half lives in DNA of N7-ethylguanine and N3-ethyladenine show only minor differences for both types of tissues, the mutagenic ethylation product 06-ethylguanine is removed from brain DNA very much more slowly than from the DNA of other tissues. Together with their high rate of DNA replication during the perinatal age, the incapacity of rat brain cells for enzymatic elimination of 06-alkylguanine from their DNA could account for an increased probability of neoplastic conversion, and hence for the NS specificity of ENU in the rat

Nervous-system-specific carcinogenesis by ethylnitrosourea in the rat: molecular and cellular aspects

A lead in the search for cellular determinants favoring neoplastic transformation may be provided by the pronounced tissue specificity of the oncogenic effect of certain carcinogens which do not require enzymatic metabolic activation, i.e., in cases where this specificity cannot be due to tissue differences in the activity of enzymes involved in the formation of the ultimate reactants. A carcinogen that fulfills this condition is the ethylating agent N-ethyl-N-nitrosourea (EtNU). Alkylation of nucleic acid constituents by N-nitroso compounds in relation to mutagenesis and carcinogenesis has received considerable attention recently

Role of CYP1B1 in PAH-DNA adduct formation and breast cancer risk

This study investigated the hypothesis that increased exposure to polycyclic aromatic hydrocarbons (PAHs) increases breast cancer risk. PAHs are products of incomplete burning of organic matter and are present in cigarette smoke, ambient air, drinking water, and diet. PAHs require metabolic transformation to bind to DNA, causing DNA adducts, which can lead to mutations and are thought to be an important pre-cancer marker. In breast tissue, PAHs appear to be metabolized to their cancer-causing form primarily by the cytochrome P450 enzyme CYP1B1. Because the genotoxic impact of PAH depends on their metabolism, we hypothesized that high CYP1B1 enzyme levels result in increased formation of PAH-DNA adducts in breast tissue, leading to increased development of breast cancer. We have investigated molecular mechanisms of the relationship between PAH exposure, CYP1B1 expression and breast cancer risk in a clinic-based case-control study. We collected histologically normal breast tissue from 56 women (43 cases and 13 controls) undergoing breast surgery and analyzed these specimens for CYP1B1 genotype, PAH-DNA adducts and CYP1B1 gene expression. We did not detect any difference in aromatic DNA adduct levels of cases and controls, only between smokers and non-smokers. CYP1B1 transcript levels were slightly lower in controls than cases, but the difference was not statistically significant. We found no correlation between the levels of CYP1B1 expression and DNA adducts. If CYP1B1 has any role in breast cancer etiology it might be through its metabolism of estrogen rather than its metabolism of PAHs. However, due to the lack of statistical power these results should be interpreted with caution

CYP1B1 expression, a potential risk factor for breast cancer

CYP1B1 expression in non-tumor breast tissue from breast cancer patients and cancer-free individuals was determined to test the hypothesis that high CYP1B1 expression is a risk factor for breast cancer. Large interindividual variations in CYP1B1 expression were found with CYP1B1 levels notably higher in breast cancer patients than cancer-free individuals. The results indicate that CYP1B1 might play a role in breast cancer either through increased PAH activation or through metabolism of endogenous estrogen to a carcinogenic derivative

CYP1B1 expression, a potential risk factor for breast cancer

CYP1B1 expression in non-tumor breast tissue from breast cancer patients and cancer-free individuals was determined to test the hypothesis that high CYP1B1 expression is a risk factor for breast cancer. Large interindividual variations in CYP1B1 expression were found with CYP1B1 levels notably higher in breast cancer patients than cancer-free individuals. The results indicate that CYP1B1 might play a role in breast cancer either through increased PAH activation or through metabolism of endogenous estrogen to a carcinogenic derivative