Biomedical subjects

E Siles

Publications and source records attributed to E Siles.

7 recordsLinked to original sources

PubMed1998

DNA damage and prediction of radiation response in lymphocytes and epidermal skin human cells.

The success of radiotherapy in eradicating tumours depends on the total radiation dose, but what limits this dose is the tolerance of the normal tissues within the treatment volume. Studies involving fibroblast survival have demonstrated the theoretical feasibility of a predictive assay of radiation sensitivity, but such an assay is still far from clinical application. Using pulsed-field gel electrophoresis (PFGE), we have quantified the initial "apparent" number of DNA double-strand breaks (dsb) induced by the radiation as an alternative measure of sensitivity in 2 different normal cell types from the same patients, epidermal skin cells and lymphocytes. We found significant inter-individual variation in the measured dsb (1-5 dsb/Gy/DNA unit). We also found a linear correlation between molecular damage in lymphocytes and skin samples from the same patient (slope = 0.83; r = 0.694; p = 0.0001). These results suggest that the initial number of dsb could be used as an indicator of the in vivo response to radiation.

Aged↗

PubMed1997

A comparison of p53 and p16 expression in human tumor cells treated with hyperthermia or ionizing radiation.

To assess the potential relationship between p53 and p16 proteins in the cellular response to stress, we have examined the levels of these proteins in a series of human tumor cell lines after treatment with either ionizing radiation or hyperthermia. We found that cells with abnormal radiation-induced G1 arrest (non-functional p53) had significantly higher constitutive levels of p16 than cells showing a normal G1 arrest (functional p53). Time-course experiments were done to test the effect of gamma-irradiation on intracellular levels of p16. The pattern of changes in p16 response was similar in all cell lines studied, and p16 expression was not related to cellular sensitivity to radiation or to the level of p53 induction after treatment. We also provide evidence that short-term exposure to high temperature causes p53 accumulation. Hyperthermia-induced p53 accumulation was greatest in those cells exhibiting the highest radiation-induced p53 accumulation, suggesting a possible relationship between p53 induction after these 2 different stresses. p16 synthesis was also induced in different cell lines after heat treatment, and this response was independent of p53 functionality. When we compared the level of p16 expression with the extent of G0/G1 arrest induced by heat, a linear correlation was found, raising the possibility that p16 may be involved in the control of cell cycle progression in response to heat treatment.

Carrier Proteins↗

PubMed1996

Relationship between p53 status and radiosensitivity in human tumour cell lines.

We examined the relationship between p53 levels before and after irradiation, radiation-induced cell cycle delays, apoptotic cell death and radiosensitivity in a panel of eight human tumour cell lines. The cell lines differed widely in their clonogenic survival after radiation, (surviving fraction at 2 Gy: SF2=0.18-0.82). Constitutive p53 protein levels varied from 2.2 +/- 0.4 to 6.3 +/- 0.3 optical density units (OD) per 10(6) cells. p53 after irradiation (6 Gy) also varied between the cell lines, ranging from no induction to a 1.6-fold increase in p53 levels 4 h after treatment. p53 function was also assessed by G1 cell cycle arrest after irradiation. The cellular response to radiation, measured as G0/G1 arrest, and the induction of apoptosis were in good agreement. However, a trace amount of DNA ladder formation was found in two cell lines lacking G1 arrest. Overall cellular radiosensitivity correlated well with the level of radiation-induced G1 arrest (correlation coefficient r=0.856; P=0.0067), with p53 constitutive levels (r=0.874, P=0.0046), and with p53 protein fold induction (r=-0.882, P=0.0038). Our data suggest that (1) the constitutive p53 level, (2) G1 arrest after irradiation, or (3) the p53 protein response to radiation may be good predictive tests for radiosensitivity in some cell types.

Apoptosis↗

PubMed1996

Radiosensitivity of human breast cancer cell lines of different hormonal responsiveness. Modulatory effects of oestradiol.

Treatments which inhibit or retard progression of the cell through the cell cycle have been reported to reduce the effectiveness of ionizing radiation by increasing cellular radioresistance. We studied cellular radiosensitivity and radiation-induced DNA damage (double-strand break, dsb) in both hormone-sensitive and non-sensitive human breast cancer cell lines. After 72h of culture in an oestradiol-deprived medium, MCF-7 BUS and T47D B8 breast cancer cells showed a significant delay in growth, whereas no effect was seen in EVSA-T cell line. In oestradiol-free medium, MGF-7 BUS cells were arrested mainly in G(zero)/G1 phase (85-90% in G(zero)/G1, 5-7% in S, and 6-8% in G2/M). The growth-delayed MCF-7 BUS cells showed reduced radiosensitivity (survival fraction at 2 Gy, SF2 = 63%; initial DNA damage 1.00 dsb/Gy/DNA unit) in comparison with proliferating cells (SF2 = 33%, initial DNA damage 2.70 dsb/Gy/DNA unit). The radio-protective effect of oestrogen deprivation was abolished by rescuing MCF-7 cells with oestrogen-containing medium. At 24h after rescue, MCF-7 BUS cells reached a cell cycle distribution close to that found under standard culture conditions and their radiosensitivity was correspondingly increased (SF2 = 40%, DNA damage = 2.52 dsb/Gy/DNA unit). Our findings indicate that: (1) sensitivity to radiation and the proportion of proliferating cells are probably related, and (2) differences in radiosensitivity reflect differences in radiation-induced DNA damage.

Breast Neoplasms↗

PubMed1995

Relationship between doxorubicin cell sensitivity, drug-induced DNA double-strand breaks, glutathione content and P-glycoprotein in mammalian tumor cells.

We have measured the cytotoxic effect of 1 h exposure to doxorubicin (DOX) on a panel of tumor cell lines. Cellular effects were measured by monolayer colony-forming assay and a colorimetric cytotoxicity assay. As parameters of chemosensitivity we used two different end-points: the dose of DOX that reduces to 50% the number of colonies (ID50) and the dose of DOX that reduces the final optical density to 50% of the control value (IC50). There was a significant correlation between both chemosensitivity indices (r = 0.886, p = 0.0034). DOX-induced DNA double-strand breaks (dsb) were evaluated using pulsed-field gel electrophoresis (PFGE) and compared with cellular effects, P-glycoprotein expression (P-170) and intracellular glutathione (GSH) levels. Our results showed a relationship between the slope of DNA dsb dose-response curves and the percentage of cells that express P-170 (r = -0.957, p = 0.0002). Our study also detects a positive relationship between cellular chemosensitivity parameters and GSH content [ID50 versus GSH (r = 0.794, p = 0.0186), IC50 versus GSH (r = 0.790, p = 0.0198)] in our panel of cell lines.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1995

Subclinical signs of kidney dysfunction following short exposure to silica in the absence of silicosis.

Signs of kidney dysfunction detectable in urinary protein excretion were searched for in a group of 86 silica-exposed workers who were compared to 86 control subjects matched for age, smoking status and body mass index. No worker had any clinical, spirometric or radiographic sign of silicosis, and exposure duration averaged 15.2 months (range: 11-20). An increase in the urinary excretion of albumin, transferrin, retinol-binding protein and N-acetyl-beta-D-glucosaminidase was found in the exposed group, and the prevalences of pathological values were also elevated in this group. By contrast, both groups had similar serum levels of creatinine and beta 2-microglobulin. These results strongly suggest that occupational exposure to silica may lead to subclinical renal effects after less than 2 years and in the absence of silicosis.

Adult↗

PubMed1994

Early decrease of serum Clara cell protein in silica-exposed workers.

Clara cell protein (CC16) is a 16 kDa protein secreted by nonciliated cells of the tracheobronchial tree; it has recently been proposed as a peripheral marker of respiratory epithelial injury. The concentration of CC16 was measured in the serum and, when available, in the sputum of 86 miners exposed to silica and of 86 control subjects matched for age, body mass index and smoking status (26 lifelong nonsmokers and 60 current smokers in both groups). Workers were exposed to silica-rich dust in a quarry for 15.2 months on average. No difference between exposed and control workers could be detected with regard to respiratory symptoms, chest radiographs or lung function tests. By contrast, the concentration of CC16 in serum was decreased in silica-exposed workers (geometric mean 12.3 micrograms.l-1) compared to controls (16.3 micrograms.l-1). The decrease was found both in lifelong nonsmokers (14.7 vs 21.9) and current smokers (11.3 vs 14.5). In the latter, tobacco smoking caused a decrease of serum CC16 that was additional to that associated with silica exposure. The determination of CC16 in sputum samples, judged to be reliable on the basis of the CC16/alpha-amylase concentration ratio (mostly from smokers), also revealed a reduction of CC16 following silica exposure (46.2 vs 106 mg.l-1). We conclude that alterations in the serum concentrations of CC16 probably reflect very early toxic effects of silica particles on the respiratory epithelium. This reinforces the view that serum CC16 is a sensitive marker, which might improve our ability to detect exposure to chemicals potentially harmful to the respiratory tract.

Adult↗