Ivo Hrazdira

Ultrasonically induced alterations of cultured tumour cells

OBJECTIVE The aim of this study was to establish: (i) which phase of the cell cycle is most sensitive to ultrasonic action; and (ii) whether and in which way ultrasound can influence components of the cytoskeleton. METHODS HeLa cell monolayers grown on glass cover-slips in DEM medium were used in all experiments. For proliferation studies, the cell monolayers were trypsinized and the cells were resuspended in fresh medium. The structure of the cytoskeleton was studied by means of the indirect immunofluorescence method. The cells were sonicated by a cw ultrasound of 0.8 MHz at low SA intensities (50, 100 and 500 mW/cm2) for 5 and 10 min. RESULTS The analysis of proliferation demonstrated that cells were most sensitive when undergoing M- and S-phases of the cell cycle. The ultrasonically induced disassembly of cytoskeleton components was most marked in microtubules and microfilaments due to depolymerization of basic proteins (tubulin and actin). The reaction of intermediate filaments was distinctly weaker. CONCLUSIONS In-vitro treatment of tumour cells with low intensity ultrasound results in partial inhibition of proliferation as well as in partial disassembly of all components of the cytoskeleton. Ultrasonically induced changes of the cytoskeleton seem to be non-specific and temporary.

Effect of ultrasound on DNA synthesis in tumor cells

The objective of this study was to consider the influence of continuous-wave (CW) ultrasound (800 kHz) on DNA synthesis in Ehrlich ascitic tumor cells in vitro. The 10-min irradiation was carried out in rotating polyethylene tubes (5 rpm). Incorporation of 3H-thymidine was employed to detect DNA synthesis. Replication and unscheduled (repair) DNA synthesis were distinguished by means of hydroxyurea treatment before insonation. It was established that the use of 0.5 W/cm2 and 1.0 W/cm2 (ISA) CW ultrasound interfered with DNA synthesis showing up as inhibition. The inhibition became most noticeable when starting insonation in a sample cooled to 5 degrees C. During insonation, the sample temperature was allowed to increase to the ambient (water bath) temperature of 37 degrees C. However, at an ultrasound intensity (SA) of 0.1 W/cm2 such inhibition was not proven, but a moderate stimulation of DNA synthesis was demonstrated. Changes in DNA synthesis after insonation were found to be transient because of a lack of stimulatory and/or inhibitory ultrasound effects after 1-h incubation at 37 degrees C. In our experimental conditions no effects on DNA synthesis due to the effects of the insonated medium on the cells were discernible. No statistically significant changes in unscheduled DNA synthesis were observed. Possible action mechanisms are discussed.