The effect of adriamycin on HeLa S-3 cells grown in monolayers was studied. Adriamycin is a newantitumor antibiotic of the anthracycline group isolated from cultures of Streptomyces peucetius var. caesiusly. Its chemical structure is very close to that of daunomycin from which it differs only in having a hydroxyl group substituted for a hydrogen atom of the acetyl radical. The effect of adriamycin on several experimental tumor systems has recently been reported from our laboratory2). The present studies were undertaken to show the cytological effects of adriamycin on HeLa S-3 cells in vitro. Furthermore, studies were made of its action on the biosynthesis of macromolecules by HeLa S-3 cells. Thymidine-6-3H with specific activity of 5.0 Ci/mmole, uridine-5-3H with specific activity of 15Ci/mmole and leucine-4,5-3H with specific activity of 32 Ci/mmole were purchased from Daiichi Pure Chemicals Co., Ltd. Materials and Methods ChemicalsTissue culture procedures : Clone S-3 of HeLa cells was used in these experiments.This clone was cultured in Rouxbottles containing 50 ml of growth mediumwhich consisted of YLE basal medium (Earle's balanced salt solution containing 0.1 % yeastolate, 0.5% lactalbumin hydrolysate and 0.ll % sodium bicarbonate) supplemented with 10 % calf serum. For observation of the effect of adriamycin on the growth of cells, a fully grown culture was treated with 0.1 % trypsin solution, and the released cells were suspended in the above mediumto approximately 2X105cells per ml. Oneml of the suspension was inoculated into each culture tube and incubated at 37°C for 24 hours in a slanted position. All experiments on asynchronous populations were performed on cells in the logarithmic phase of growth. Adriamycin at desired concentration was added on 0 day. Three tubes
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