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1.
Exp Cell Res ; 319(20): 3094-103, 2013 Dec 10.
Article in English | MEDLINE | ID: mdl-23973668

ABSTRACT

The mortality of patients with solid tumors is mostly due to metastasis that relies on the interplay between migration and proliferation. The "go or grow" hypothesis postulates that migration and proliferation spatiotemporally excludes each other. We evaluated this hypothesis on 35 cell lines (12 mesothelioma, 13 melanoma and 10 lung cancer) on both the individual cell and population levels. Following three-day-long videomicroscopy, migration, proliferation and cytokinesis-length were quantified. We found a significantly higher migration in mesothelioma cells compared to melanoma and lung cancer while tumor types did not differ in mean proliferation or duration of cytokinesis. Strikingly, we found in melanoma and lung cancer a significant positive correlation between mean proliferation and migration. Furthermore, non-dividing melanoma and lung cancer cells displayed slower migration. In contrast, in mesothelioma there were no such correlations. Interestingly, negative correlation was found between cytokinesis-length and migration in melanoma. FAK activation was higher in melanoma cells with high motility. We demonstrate that the cancer cells studied do not defer proliferation for migration. Of note, tumor cells from various organ systems may differently regulate migration and proliferation. Furthermore, our data is in line with the observation of pathologists that highly proliferative tumors are often highly invasive.


Subject(s)
Cell Movement , Cytokinesis , Models, Biological , Neoplasms/pathology , Cell Proliferation , Humans , Tumor Cells, Cultured
2.
J Photochem Photobiol B ; 120: 120-9, 2013 Mar 05.
Article in English | MEDLINE | ID: mdl-23375215

ABSTRACT

Conventional photodynamic diagnosis (PDD) and therapy (PDT) makes use of photosensitizers that are excited by continuous light irradiation of specific wavelengths. In the case of PDT, the overdose of continuous excitation may lead to an expansion of necrosis in cancer cells or morbidity in healthy surroundings. The present study involves 5-h fluorescence imaging of living human lung epithelial carcinoma cells (A549) in the presence of a novel photosensitizer, PVP-Hypericin (PVP: polyvinylpyrrolidone) to optimize the excitation light doses for PDD and PDT. A number of time-lapse imaging experiments were performed using a low-power blue LED operating in either continuous or pulsed mode. The irradiances I(*) were 1.59, 6.34 and 14.27mW/cm(2), the pulse lengths L being 0.127, 1.29, 13, 54.5, 131 and 60,000ms. Then, the relation between irradiance, various exposure times, photobleaching and phototoxicity of PVP-Hyperycin was investigated. Results showed a nonlinear relationship between the amounts of excitation dose, cell viability and toxicity. For all experimental I(*), minimal phototoxicity and photobleaching was detected when cells were exposed to brief pulses of light (L⩽13ms). On the other hand, pulsed excitation with I(*)=14.27mW/cm(2) and L=131ms induced high percentages of apoptosis comparable to the long exposures of L=60,000ms and the continuous excitation. Thus, replacement of continuous excitation by a pulsed method seems applicable for PDT.


Subject(s)
Diagnostic Imaging/methods , Light , Perylene/analogs & derivatives , Photochemotherapy/methods , Povidone/therapeutic use , Anthracenes , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Cell Survival/drug effects , Cell Survival/radiation effects , Dose-Response Relationship, Radiation , Fluoresceins/metabolism , Humans , Perylene/pharmacology , Perylene/therapeutic use , Photobleaching , Photosensitizing Agents/pharmacology , Photosensitizing Agents/therapeutic use , Povidone/pharmacology , Succinimides/metabolism , Time Factors
3.
Arch Toxicol ; 86(5): 805-13, 2012 May.
Article in English | MEDLINE | ID: mdl-22331240

ABSTRACT

Glyphosate (G) is the largest selling herbicide worldwide; the most common formulations (Roundup, R) contain polyoxyethyleneamine as main surfactant. Recent findings indicate that G exposure may cause DNA damage and cancer in humans. Aim of this investigation was to study the cytotoxic and genotoxic properties of G and R (UltraMax) in a buccal epithelial cell line (TR146), as workers are exposed via inhalation to the herbicide. R induced acute cytotoxic effects at concentrations > 40 mg/l after 20 min, which were due to membrane damage and impairment of mitochondrial functions. With G, increased release of extracellular lactate dehydrogenase indicative for membrane damage was observed at doses > 80 mg/l. Both G and R induced DNA migration in single-cell gel electrophoresis assays at doses > 20 mg/l. Furthermore, an increase of nuclear aberrations that reflect DNA damage was observed. The frequencies of micronuclei and nuclear buds were elevated after 20-min exposure to 10-20 mg/l, while nucleoplasmatic bridges were only enhanced by R at the highest dose (20 mg/l). R was under all conditions more active than its active principle (G). Comparisons with results of earlier studies with lymphocytes and cells from internal organs indicate that epithelial cells are more susceptible to the cytotoxic and DNA-damaging properties of the herbicide and its formulation. Since we found genotoxic effects after short exposure to concentrations that correspond to a 450-fold dilution of spraying used in agriculture, our findings indicate that inhalation may cause DNA damage in exposed individuals.


Subject(s)
DNA Damage , Epithelial Cells/drug effects , Glycine/analogs & derivatives , Herbicides/toxicity , Cell Line , Cheek , Dose-Response Relationship, Drug , Glycine/toxicity , Humans , L-Lactate Dehydrogenase/metabolism , Mitochondria/drug effects , Occupational Exposure , Toxicity Tests , Glyphosate
4.
Electrophoresis ; 30(8): 1306-28, 2009 Apr.
Article in English | MEDLINE | ID: mdl-19382132

ABSTRACT

Interpretation of proteome profiling experiments largely relies on comparative analyses. False-positive identifications may cause fatal misinterpretation of data. On the other hand, proteome analysis may also suffer from false negatives, when proteins that are actually present are not detected. This circumstance may be as fatal as false-positive identifications and was hardly considered until now. Appropriate positive controls would facilitate quality assessment of proteome profiling experiments. Based on cell biology knowledge, our aim was to generate a list of commonly expressed proteins, which may serve as positive control. Following a pragmatic experimental strategy, we compared the cytoplasmic fractions of four largely differing kinds of cells, which were human DCs, endothelial cells, fibroblasts and keratinocytes. Proteome profiling was performed by 2D-PAGE in addition to shotgun analysis. By shotgun analysis, 665 proteins were identified, which occurred in each of the four cells types; 360 proteins of those were also detectable in the corresponding 2-D gels. We consider these proteins as common proteins. All shotgun analysis data, including mass fragmentation spectra of the corresponding peptides, are accessible via the proteomics identification database (http://www.ebi.ac.uk/pride). As expected, most of the common proteins could be clearly assigned to at least one of the following functional categories: chaperones, cytoskeleton, energy metabolism, redox regulation, nucleic acid processing, protein turnover, membrane transport, protein synthesis and signaling. We suggest that the present data may prove helpful for data assessment, quality control and interpretation of a large variety of experiments based on proteome profiling.


Subject(s)
Electrophoresis, Gel, Two-Dimensional , Peptide Mapping , Proteins/analysis , Proteomics/methods , Databases, Protein , Dendritic Cells/metabolism , Endothelial Cells/metabolism , Fibroblasts/metabolism , Gene Expression , Humans , Keratinocytes/metabolism
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