Distinct effects of ionizing radiation on in vivo murine kidney and brain normal tissue gene expression.

PURPOSE: There is a growing awareness that radiation-induced normal tissue injury in late-responding organs, such as the brain, kidney, and lung, involves complex and dynamic responses between multiple cell types that not only lead to targeted cell death but also acute and chronic alterations in cell function. The specific genes involved in the acute and chronic responses of these late-responding normal tissues remain ill defined; understanding these changes is critical to understanding the mechanism of organ damage. As such, the aim of the present study was to identify candidate genes involved in the development of radiation injury in the murine kidney and brain using microarray analysis. EXPERIMENTAL DESIGN: A multimodality experimental approach combined with a comprehensive expression analysis was done to determine changes in normal murine tissue gene expression at 8 and 24 hours after irradiation. RESULTS: A comparison of the gene expression patterns in normal mouse kidney and brain was strikingly different. This observation was surprising because it has been long assumed that the changes in irradiation-induced gene expression in normal tissues are preprogrammed genetic changes that are not affected by tissue-specific origin. CONCLUSIONS: This study shows the potential of microarray analysis to identify gene expression changes in irradiated normal tissue cells and suggests how normal cells respond to the damaging effects of ionizing radiation is complex and markedly different in cells of differing origin.

Integrins may serve as mechanical transducers for low-frequency electric fields.

A hypothesis is presented that a transduction mechanism for low frequency electric fields of physiological strength ( approximately 1 V/cm) is the same as that for sinusoidal fluid shear stresses, the force exerted on an integrin. Simple calculations show that the forces exerted on a model integrin by transverse electric fields and fluid shears that produce cellular effects are comparable in magnitude, about 1 fN. The electric force is provided by the interaction of the surface charges on the integrin with the tangential component of the applied field. The mechanical shear force is the transverse fluid drag force exerted on the cylindrical surface of the integrin. Either force is coupled mechanically to the actin cortex within the cell. The mechanical network which exists within a cell and connects a cell to its surroundings would then be directly coupled to an applied electric field. The fundamental transduction mechanism for some electric field effects may then be ultimately mechanical in nature.

Particle irradiation suppresses metastatic potential of cancer cells.

Particle radiotherapy such as proton and carbon ion has been producing promising clinical results worldwide. The purpose of this study was to compare metastatic capabilities of malignant tumor cells after irradiation with photon, proton, and carbon ion beams to clarify their ion beam-specific biological effects. We examined the biological properties of highly aggressive HT1080 human fibrosarcoma cells to assess their metastatic processes in terms of cell adhesion capability to extracellular matrix, expression of integrins, cell migration, cell invasive capability, and matrix metalloproteinase-2 activity in vitro. We then assessed the metastatic capabilities of LM8 mouse osteosarcoma irradiated with carbon ion or photon beam in the syngeneic mice. Both proton and carbon ion irradiation decreased cell migration and invasion in a dose-dependent manner and strongly inhibited matrix metalloproteinase-2 activity. On the other hand, lower X-ray irradiation promoted cell migration and invasion concomitant with up-regulation of alphaVbeta3 integrin. For cancer cells treated with carbon ion irradiation, the number of pulmonary metastasis was decreased significantly in vivo. These findings suggest that particle irradiation suppresses metastatic potential even at lower dose, whereas photon irradiation promotes cell migration and invasive capabilities at lower dose level, and provide preclinical evidence that ion beam radiotherapy may be superior to conventional photon beam therapy in possible preventive effects on metastases of irradiated malignant tumor cells.

Integrin-mediated targeting of drug delivery to irradiated tumor blood vessels.

The objective of this study was to target drug delivery to radiation-induced neoantigens, which include activated receptors within the tumor vasculature. These responses include posttranslational changes in pre-existing proteins, which can be discovered by phage-displayed peptide libraries administered to mice bearing irradiated tumors. Phage-displayed peptides recovered from irradiated tumors included the amino acid sequence RGDGSSV. This peptide binds to integrins within the tumor microvasculature. Immunohistochemical staining of irradiated tumors showed accumulation of fibrinogen receptor alpha(2b)beta(3) integrin. We studied tumor targeting efficiency of ligands to radiation-induced alpha(2b)beta(3). Radiopharmaceuticals were localized to irradiated tumors by use of alpha(2b)beta(3) ligands conjugated to nanoparticles and liposomes. Fibrinogen-conjugated nanoparticles bind to the radiation-activated receptor, obliterate tumor blood flow, and significantly increase regression and growth delay in irradiated tumors. Radiation-guided drug delivery to tumor blood vessels is a novel paradigm for targeted drug delivery.

Irradiation induces increase of adhesion molecules and accumulation of beta2-integrin-expressing cells in humans.

PURPOSE: The purpose of our investigation was to describe the dose- and time-dependent histomorphologic alterations of the irradiated tissue, the composition of the infiltrate, and the expression patterns of various adhesion molecules. METHODS AND MATERIALS: We analyzed immunohistochemically alterations in oral mucosa in 13 head and neck cancer patients before radiotherapy and with 30 Gy and 60 Gy. All had oral mucosa irradiation, with a final dose of 60 Gy using conventional fractionation. Snap-frozen specimens were stained using the indirect immunperoxidase technique. Histomorphology was studied in paraffin-embedded sections. In addition, we determined the clinical degree of oral mucositis. RESULTS: Histomorphologic evaluation showed no vascular damage. Irradiation caused a steep increase of beta2-integrin-bearing cells (p < 0.01), whereas the percentage of beta1-integrin-positive cells remained at low levels. Additionally we found an increase in the expression of endothelial intercellular adhesion molecule-1 (ICAM-1) (p < 0.01) and E-selectin (p < 0.05), while endothelial vascular cell adhesion molecule-1 (VCAM-1) expression remained at very low levels. CONCLUSION: Our findings indicate that in radiation-induced oral mucositis there is no marked vascular damage until the end of radiotherapy. For recruitment of leukocytes, beta2 is more involved than beta1. Pharmaceuticals that block leukocyte adhesion to E-selectin or ICAM-1 may prevent radiation-mediated inflammation in oral mucosa.

Preferential repair in Saccharomyces cerevisiae rad mutants after induction of interstrand cross-links by 8-methoxypsoralen plus UVA.

The gene specific induction and the incision step of the removal of 8-methoxypsoralen (8-MOP) plus UVA-induced interstrand cross-links (ICL) was measured in repair mutants of Saccharomyces cerevisiae. Events were examined at the MAT alpha and HML alpha loci in mutants deficient in the repair of ICL, namely rad1, rad2 delta, rad52, pso2 and the rad16 mutant which is impaired in the removal of UV-induced pyrimidine dimers from the silent HML alpha locus. Previously, we observed in a wild-type strain (K107) preferential repair concerning the incision of 8-MOP photo-induced ICL. The present study indicates that the two mutants rad1 and rad2 delta show no repair in either locus, due presumably to their deficiency in the incision step of ICL repair. The rad52 mutant which is defective in recombination, is proficient in the preferential incision of ICL at the MAT alpha locus versus the HML alpha locus. The same is true for the pso2 mutant which also lacks the ability to perform complete repair of ICL. The rad16 mutant is unable to repair ICL in the silent locus HML alpha but is proficient in repair (i.e. the incision of ICL) in the transcriptionally active MAT alpha locus.

Electro-magnetic fields in the home environment (color TV, computer monitor, microwave oven, cellular phone, etc) as potential contributing factors for the induction of oncogen C-fos Ab1, oncogen C-fos Ab2, integrin alpha 5 beta 1 and development of cancer, as well as effects of microwave on amino acid composition of food and living human brain.

The effects, on normal human subjects, of 3 minutes exposure to electro-magnetic fields (EMFs) emitted from: A) personal computers, B) color television sets, or C) microwave-ovens, or cellular phones were compared by placing the same large sheet of aluminum foil with a square hole or rectangular band-shaped hole at the chest level (or at the side of the head with the cellular phone), with or without grounding the aluminum foil, using the Bi-Digital O-Ring Test Dysfunction Localization and Molecular Identification Methods with cancer related substances (i.e., Oncogen C-fos Ab2 and mercury in the cell nucleus, Integrin alpha 5 beta 1 in the cell & nuclear membranes, and disappearance of Acetylcholine) as reference control substances. All the above sources of the EMFs not only induced the following various transitional abnormalities on the EMF entry area, but also induced similar abnormalities at the EMF exit area on the back (where the abnormality was found in the same shape as exposed EMF entry area, and the effect lasted for a shorter time than the entry point of the EMF): A) Exposure of the body at about 50 cm from the monitor of some of the typical personal computers resulted in: A1) decrease in Acetylcholine; A2) appearance of circulatory disturbance with the appearance of Thromboxane B2; A3) short-lasting appearance of Oncogen C-fos Ab2; A4) short-lasting appearance of Oncogen C-fos Ab1, though it lasted longer than C-fos Ab2; A5) no appearance of Integrin alpha 5 beta 1. B) part of the chest was exposed at a distance between 1 meter and up to 3 meters from a color television sized anywhere from 13'' to 21'', resulting in: B1) decrease in Acetylcholine; B2) appearance of circulatory disturbance with the appearance of Thromboxane B2; B3) short-lasting appearance of Oncogen C-fos Ab2; B4) short-lasting appearance of Oncogen C-fos Ab1, though it lasted longer than C-fos Ab2; B5) very short-lasting appearance of Integrin alpha 5 beta 1. C) When body was exposed, at a distance of 0.5m-2 meters, to microwaves emitted as leakage from a small microwave oven (about 2.45 GHz with 450 Watt output), the effects usually lasted about 2 to 3 times the exposure time at the exposed area and 1.6 to 2 times the exposure time at the back of the body at the EMF exit area.(ABSTRACT TRUNCATED AT 400 WORDS)