Abstract
It is well known that a number of -SH and -SS containing substances afford a certain measure of protection against radiation effects in many biological systems, and it is conceivable that inherent -SH levels in Ehrlich ascites tumour (ELD)cells may be of decisive improtance with respect to the development of cellular radiation injury. So far, little effort has been directed to elucidate the changes in levels of different -SH and -SS groups in ELD cells when the tumour-bearing whole animal was subjected to the sublethal dose of X-irradiation. The present study was designed to bring some lights in the possible changes of and relationship between various sulfhydryl levels, such as P-SH, NP-SH and NP-SS, as well as the content of protein and cell volume of ELD cells, after subjecting the ELD mice to 1,200 r of X-irradiation. The animals used in this experiment were all mixed bred mice of $20{\sim}25\;gm$ in body weight (approximately 2 months old) irrespective of sex. 12 mice in one experiment were inoculated intraperitoneally with 0.2 ml of ascites tumour cells $(2{\times}10^6\;cells)$, and on the 7th day of the tumour growth, they were X-irradiated with 1,200 r, using the conventional X-ray machine under the following conditions: 200 Kv at 15 mA, 0.5 mm Cu filter, target-skin distance: 50 cm. Radiation dose was measured with the the Philip integrating dosimeter. At 24, 36, 48 and 60 hours after the X-irradiation, the mice were killed by cervical dislocation, and the tumours were taken out. Freshly withdrawn ascites tumours were placed in ice, and immediately the cell concentration was measured with the Coulter Cell Counter (Model B), and the hematocrit of the tumour cells were also determined. Cell volume was thus calculated by the cell concentration and hematocrit value. P-SH content of ELD cells was measured potentiometrically according to the method of Calcutt & Doxey, and NP-SH and NP-SS contents were measured spectrophotometrically by the method described by Ellman. Protein content of ELD cells was determined with the Folin phenol reagent by Lowry et al. Altogether, 48 experimental mice were used, and 12 mice with the only exception of X-irradiation were used as the control. Results obtained indicate that the contents of all the cellular sulfhydryl groups as well as cell volume and protein content of the ELD cells increase significantly as time progresses after the sub-lethal X-ray dose of 1,200 r was given and that all the increase is in a lineal fashion. The regression lines of the relative values, (i. e., taking each control value as 1) of all the values obtained, and the regression lines of cell volume, protein and NP-SH are identical, whereas those of NP-SS and P-SH appear to be widely seperated. However, the difference of those two lines (NP-SS & P-SH) were found to be not significant statistically (p>0.05). Therefore, it can be concluded from the above results that all the values examined increase in a lineal fashion with no statistically significant difference among them. Also, with the radiation dose of 1,200 r, the ELD cell becomes enlarged and swollen progressively up to 60 hours post-irradiation and it becomes more than two times of the original normal size at 60 hours after the irradiation, and up to this stage, it seems apparent that the cell division has been slow due to the X-irradiation applied in this experiment. It is well understandable that the contents of NP-SH, NP-SS, P-SH and protein of the ELD cells increase in parallel with the increase of the cell volume by the X-ray does used, but it also seems interesting to note that all the cellular substances tested show no appreciable difference in the pattern of increase.