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Journal of Cell and Molecular Biology 1: 87-91, 2002. Golden Horn University, Printed in Turkey. The effect of adriamycin on Ehrlich ascites tumor cells iinn vviittrroo and iinn vviivvoo Gülruh Ulako¤lu University of ‹stanbul, Faculty of Science, Department of Biology, 34459, Vezneciler, ‹stanbul, Turkey Received 15 April 2002; Accepted 25 June 2002 Abstract The effect of different doses of adriamycin (ADM), was investigated in vitro and in vivo, in Ehrlich ascites tumor (EAT) cells which developed in the peritoneal cavity of mice. In the in vivo experiments it was found that injection of ADM as 10 mg/g i.p. prolonged the survival period of mice. In the in vitro experiments, treatment of ADM in 2, 4 and 6 mg/ml doses, it was observed that cytotoxic effect dependent on time occurred in tumor cells. KKeeyy wwoorrddss:: Ehrlich ascites carcinoma, adriamycin (adriablastina), in vivo, in vitro Adriamisinin iinn vviittrroo ve iinn vviivvoo koflullarda Ehrlich ascites tümör hücrelerine etkisi Özet Adriamisinin (ADM) çeflitli dozlar›n›n farelerin periton bofllu¤unda geliflen Ehrlich ascites tümör (EAT) hücrelerine etkisi in vivo ve in vitro olarak denendi. In vivo deneylerde farelere 10 mg/g dozda i.p. olarak enjekte edilen ADM in farelerin yaflam süresini uzatt›¤› saptand›. In vitro deneylerde EAT hücreleri 2, 4 ve 6 mg/ml dozda ADM ile muamele edildi¤inde, zamana ba¤l› olarak tümör hücrelerine sitotoksik etki meydana geldi¤i gözlendi. AAnnaahhttaarr ssöözzccüükklleerr:: Ehrlich ascites karsinoma, adriamisin (adriablastina), in vivo, in vitro Introduction ADM, being an anthracycline antibiotic, has a large spectrum of antitumor activity. Clinically, it was administered to various tumors singly or with other antitumor antibiotics (Sipahio¤lu, 1981). Different results have been obtained from ADM when administered in different doses both clinically and in laboratory experiments. In an experiment performed with HeLa cells, a significant decrease in the number of surviving cells occurred depending on the dose of applied ADM (Jagetia and Nayak, 1996). Experiments on the effective mechanism of ADM to tumor cells were also done. It was reported that it had the highest impact on DNA; DNA replication, transcription and repair were effected. It was also shown that it inhibited protein synthesis (Saraswathi et al., 1997). ADM interacts with cell membrane and by affecting the processes such as lectin interaction, phospholipid’s structure and organization, fluidity and transportation of small molecules and ions in the cell membrane causes cytotoxicity (Tritton and Yee, 1982). During the application of ADM in high doses clinically, its side effects on the patients have been observed (Sipahio¤lu, 1981). In an experiment, it was observed that the administration of 20 mg/kg ADM increased the microsomal lipid peroxidation level in rats which was found to be related with toxicity induced by ADM. For this reason, when ADM is used at high doses, it was administered with drugs which decrease microsomal lipid peroxidation (Shinozawa 87
88 Gülruh Ulako¤lu et al., 1993). A group of research workers, treated their breast carcinoma, gastric carcinoma and colon carcinoma patients with a single dose of 25 mg/m 2 i.v. ADM. After peripheral blood samples were obtained serially from these patients before and after drug injection were compared. They found that ADM had a cytotoxic effect on the blood cells which was its side effect (Arinaga et al., 1986). In this study, whether ADM prolonged or not the survival periods of mice bearing EAT cells was investigated. Besides the mode of effect of ADM’s different doses on EAT cells multiplication was also examined in vitro. Material and methods EAT cells used in this study were hyperdiploid carcinoma cells, and they were maintained in our laboratory by making their transplantions every 7 days. Preparation of ADM ADM isolated from Streptomyces peucetius by Farmitalia Research Laboratories for the first time. ADM (Carlo Erba, Turkey) used in the experiments was a lyophylized and stock solution obtained by dissolving 10 mg ADM in 5 ml sterile water. From this solution 2, 4, 6 mg/ml ADM concentrations were prepared to be used in the experiments. In vivo experiments In the experiments 3 months old male albino mice of strain Balb/c weighing between 25-30 g were used. The mice were first inoculated intraperitonally with 3X10 6 EAT cells and they were separated into control and experimental groups. There are 10 mice in the control group and 29 mice in the experimental group. In the in vivo experiment were used total 39 mice. 4 days after the inoculation of EAT cells, 10 mg/g ADM was injected i.p. in a single dose to the experimental mice. The control group’s mice were injected with physiological serum. The status of all the mice were controlled every day and the death dates of both the experimental and control mice were determined. In vitro experiments In these experimental series also EAT cells were drawn from the stock mice with an injector and a cell suspension was prepared in BSS. From this suspension, 3X10 5 cells/ml were transferred into steril tubes containing medium (Eagle + 2.5% Fetal calf serum + 10 mM Hepes). After the tubes were closed with steril rubber stoppers, they were incubated in an incubator at 37°C. EAT cells were separated into experimental and control tubes, 24 hours after being taken in vitro condition. ADM in 2, 4 and 6 mg/ml concentrations were added separately into the experimental tubes (Çerçi and Ulako¤lu, 2000). At the end of 3, 5, 24, 30 and 48 hours of drug addition, cell counted were done. Both the control and experimental cells were treated with trypan blue, counted under the light microscope and the amount of dead and surviving cells were determined. According to the data obtained, by the application of paired t-test their significance at p
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