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PubMed1982

Effects of the chiral isomers of erythro- and threo-9-(2-hydroxy-3-nonyl)adenine on purine metabolism in sarcoma 180 cells.

The effects of the chiral isomers of erythro- and threo-9-(2-hydroxy-3-nonyl)adenines (EHNA and THNA) on purine metabolism in Sarcoma 180 cells have been determined. At concentrations of 10-80 microM [10- to 1000-fold greater than their Ki values with adenosine deaminase (ADA)], all isomers inhibited purine salvage and biosynthesis de novo. Although (+)-EHNA, the most potent ADA inhibitor, exerted the greatest effects, there was no direct correlation between the potency of ADA inhibition and the secondary effects on purine metabolism, e.g. (+)-EHNA is about 2-fold more inhibitory than (-)-EHNA in blocking purine base incorporation but about 250-fold more potent as an inhibitor of ADA (Ki of (+)-EHNA = 2 nM; Ki of (-)-EHNA = 500 nM [Bessodes et al., Biochem. Pharmac. 31, 879 (1982)]). All the isomers inhibited the incorporation of radiolabeled purine bases (adenine, guanine and hypoxanthine) and nucleosides (guanosine and inosine) into acid-soluble nucleotides and of glycine into 5'-phosphoribosyl-formylglycineamide. Unlike the results of Henderson et al. [Biochem. Pharmac. 26, 1967 (1977)] with Ehrlich ascites cells, the incorporation of adenosine into nucleotides was only slightly inhibited in Sarcoma 180 cells. (+)-EHNA did not inhibit the activities of 5-phosphoribosyl-1-pyrophosphate (PRPP) synthetase, purine phosphoribosyltransferases or nucleotide kinases in cell extracts. Accumulation of PRPP was inhibited only under conditions that fostered rapid synthesis.

Adenine↗

PubMed1978

Nitrogen analogues of 1,4-benzoquinones. Activities against the ascitic sarcoma 180 of mice.

Compounds having the basic structure N-(R)-substituted ring-substituted 4-iminocyclohexadienone have been synthesized and tested as antitumor agents against the ascitic sarcoma 180 tumor in Swiss mice. Among these compounds, the dimethylindoanilines [R = 4-(CH3)2NC6H4] are most stable in water at pH 7.0 and at 25 degrees C, the oximes (R = oh) are less stable, and the N-halo compounds (R = Br and Cl) are least stable. The N-halo derivatives have the highest redox potentials under the conditions used, the greatest effect against ascitic sarcoma 180 in Swiss mice, and the greatest acute toxicity when injected ip in the Swiss mice. Discriminant analysis of the results indicates that substituents with positive values of F and negative values of pi increase the antitumor activities, whereas those with positive values of sigma and R should lower the toxicity. The redox potential, a molecular parameter, is the best single variable for discriminating between the groups based on antitumor activities.

Animals↗

PubMed1986

Decreased autolysis of dead cells in adriamycin-resistant lines of the sarcoma 180 of mouse.

Adriamycin-resistant cell lines of the sarcoma 180 of mouse reveal, besides the known resistance mechanism, a decrease in cellular autolysis compared with the original line. This decrease has been demonstrated by comparative analysis of cell-free supernatants isolated from 7-day-old sensitive and resistant ascites tumors in mice. In order to demonstrate these changes we isolated and quantified the DNA, detecting higher amounts within the supernatants of resistant lines. Furthermore, unspecific DNA-cleaving activity within raw homogenates of the cells is substantially lower in the resistant lines. Residual parts of chromatin may trap the drug and in this way lower its effective concentration. However, the results may also reflect changes in enzymatic complement playing a hitherto unknown role within living cells.

Animals↗

PubMed1985

Decrease in alkaline triglyceride lipase in primary cultured hepatocytes from mice with sarcoma 180.

Primary cultured hepatocytes from normal mice and mice with Sarcoma 180 were characterized. The viability of freshly isolated hepatocytes from both sources was over 90% and the cells had a relatively stable population of DNA for a minimum of three days. After incubation with (3H)leucine, the syntheses and secretions of (3H)labeled trichloracetic acid-insoluble materials by hepatocytes from both normal and tumor-bearing mice increased similarly. However, the alkaline triglyceride lipase activity of a homogenate of freshly isolated hepatocytes from tumor-bearing mice was one-third that of cells from normal mice. The activity of hepatocytes from tumor-bearing mice increased less during culture than did the activity of cells isolated from normal mice.

Animals↗

PubMed1983

Intensification of host's immunity by squalene in sarcoma 180 bearing ICR mice.

Effect of highly purified squalene (HP-SQ) obtained from liver of a shark, Centrophorus atromarginative Garman, on antitumor activity and host's immune response was studied in sarcoma 180 bearing female ICR mice. HP-SQ was administered intraperitoneally (i.p.) to solid tumor-bearing or healthy mice at the dose of 0.1 ml/mouse/d for consecutive 10 d. The administration of HP-SQ exerted the host's resistance against challenging 5 x 10(4) cells of ascites sarcoma 180, and it resulted in marked prolongation of survival of the mice (ILS; 110%). Function of the reticuloendothelial system estimated by a carbon clearance test was enhanced by i.p. administration of HP-SQ in both the healthy and the tumor-bearing mice. Number of IgM antibody forming cells against sheep red blood cells (SRBC) in the spleen was counted 4 d after intravenous injection of 2 x 10(7) cells of SRBC. The number of the IgM antibody forming cell increased by HP-SQ administration in the tumor-bearing and the healthy mice. Delayed hypersensitivity reaction against SRBC measuring swelling of the foot pads of mice was also stimulated by the i.p. administration of the material.

Animals↗

PubMedDate not supplied

Anticancer activity of new 3-amino-pyrrolidinedione-nitrogen mustard derivatives on murine sarcoma 180.

A 2,5-pyrrolidinedione linked nitrogen mustard derivative, (R,S)3-[N,N-bis(2-chloroethyl)]-amino-1-(2'-methoxyphenyl)pyrrolidine- 2,5-dione hydrochloride (I) showed a marked antiproliferative effect on mouse Sarcoma 180. Since (I) is also active against L1210 and P388 leukaemias, its toxicity in mice was evaluated. A study of acute toxicity revealed focal liver cell necrosis. Another derivative, (R,S)3-[N,N-bis-(2-chloroethyl)]amino-1-(4'-n-butoxyphenyl)pyrrolidine- 2,5-dione dihydrate (II), which also possessed significant anticancer effect on P388 and L1210 leukaemias, was inactive against Sarcoma 180.

Animals↗

PubMed1980

Overadditive chemotherapeutic synergism of prospidine and inosine when given sequentially to the Sarcoma 180 implanted intramuscularly.

The chemotherapeutic effectiveness of prospidine monotherapy on the Sarcoma 180 implanted i.m. was compared with several prospidine/inosine combinations. The chemotherapeutic action of the combination was enhanced overadditively when inosine was given 6 h after prospidine. This particular combination caused not only a significant curative action and tumor inhibition as compared with prospidine monotherapy, but also decreased the toxic effects.

Animals↗

PubMed2000

Antitumour activity and side effects of combined treatment with chitosan and cisplatin in sarcoma 180-bearing mice.

We examined the possible modulation by chitosan of the antitumour effects and side effects of cisplatin (cis-diaminedichloroplatinum, CDDP). The study showed that CDDP had potent antitumour activity when administered orally as well as intraperitoneally. We also compared the antitumour activity and side effects of orally administered CDDP plus orally administered chitosan versus intraperitoneally administered CDDP plus orally administered chitosan in sarcoma 180-bearing mice. When CDDP (1.25 mgkg(-1) x 2 day(-1)) was intraperitoneally administered to sarcoma 180-bearing mice, myelotoxicity (the reduction of leucocyte and platelet numbers), nephrotoxicity (the increase of blood nitrogen urea level), immunotoxicity (the reduction of spleen and thymus weight) and a reduction in body weight resulted. These intraperitoneally administered CDDP-induced side effects were not prevented by oral administration of chitosan (150 mgkg(-1) x 2 day(-1) and 750 mgkg(-1) x 2 day(-1)) for 14 consecutive days. On the other hand, the side effects such as the reductions of body and spleen weights induced by orally administered CDDP (1.25 mgkg(-1) x 2 day(-1)) were prevented by the oral administration of chitosan (150 mgkg(-1) x 2 day(-1) and 750 mg kg(-1) x 2 day(-1)). From these results, we conclude that the orally administered chitosan plus CDDP might be useful for the prevention of body weight reduction and immunotoxicity (the reduction of spleen weight) induced by the orally administered CDDP without diminishing antitumour activity.

Administration, Oral↗

PubMed1982

Increased relative effectiveness of doxorubicin against slowly proliferating sarcoma 180 cells after prolonged drug exposure.

Doxorubicin (ADR) lethality was studied in Sarcoma 180 cells grown in vitro during log phase and early plateau phase. For both log phase and plateau phase cells, ADR lethality increased progressively with continuous drug exposure, and was dependent on drug concentration at every point in time. For any given level of toxicity to log phase cells, greater killing of plateau phase cells was achieved with prolonged drug exposure. The inhibition of DNA synthesis by ADR was delayed in onset, and increased gradually, both at low and high drug concentrations. The data suggest that the dependence of ADR lethality on proliferation rate and the higher therapeutic ratio in plateau phase cells with prolonged drug exposure are not directly related to the ADR effects on DNA synthesis.

Animals↗

PubMed1971

A polynucleotide segment rich in adenylic acid in the rapidly-labeled polyribosomal RNA component of mouse sarcoma 180 ascites cells.

The rapidly-labeled polyribosomal RNA component from mouse sarcoma 180 cells is retained on nitrocellulose (Millipore) membrane-filters at high ionic strength. This property is due to the presence of a polynucleotide sequence rich in adenylic acid that resists both T(1) and pancreatic RNase digestion. The resistant material shows sedimentation characteristics close to those of transfer RNA. The RNA molecules that contain this material can be separated from the rest of the polysomal RNA by differential phenol extraction with neutral and alkaline Tris buffers. Synthetic poly(A) exhibits the same behavior as the rapidly-labeled polysomal RNA with respect to Millipore binding and phenol fractionation. The characteristics of the rapidly-labeled polysomal RNA component permit its isolation free of ribosomal RNA.

Adenine Nucleotides↗

PubMed1973

In vitro translation of oogenetic messenger RNA of sea urchin eggs and picornavirus RNA with a cell-free system from sarcoma 180.

A cell-free protein-synthesizing system prepared from mouse sarcoma 180 was characterized by use of RNA from mengo virus and sea urchin egg. In the presence of exogenous mammalian transfer RNA, total sea urchin egg RNA and mengo RNA direct incorporation of [(3)H]leucine into acid-insoluble material. The system is extremely efficient in that a stimulation of 100-times over background can be obtained. Studies with formylmethionyl-transfer RNA, as well as with inhibitors of initiation, indicate that multiple initiation occurs; further, 85-90% of all chains made in vitro are subsequently released from ribosomes. An average translation time of 3.5 min was determined with messenger RNA of sea urchin egg, and product analysis indicates that high-molecular-weight products (greater than 50,000 molecular weight) are being made in vitro. Sequences of sea urchin egg RNA containing poly(A) act as messenger RNA.

Animals↗

PubMed1998

Hepatocyte growth factor/scatter factor activates the apoptosis signaling pathway by increasing caspase-3 activity in sarcoma 180 cells.

Hepatocyte growth factor, which is now known to be the same protein as scatter factor, induced oligonucleosomal fragmentation of nuclear DNA of Sarcoma 180 cells and increased the activity of caspase-3, a key component in control of the apoptotic cell death pathway to about 2.6 times that in control cells on 48 hr incubation, but did not increase the activity of caspase-1. Both HGF-induced DNA fragmentation and caspase-3 activity were completely inhibited by co-incubation with an inhibitor of caspase-3, Ac-DEVD-H. In contrast, HGF did not affect the expression of the apoptosis suppressors Bcl-2 and Bcl-x. These results indicate that HGF activates the apoptosis signaling pathway by increasing caspase-3 activity in Sarcoma 180 cells.

Animals↗

PubMed1976

Potentiation by guanine nucleosides of the growth-inhibitory effects of adenosine analogs on L1210 and sarcoma 180 cells in culture.

The growth-inhibitory effect of 6-methylmercaptopurine riboside (MMPR) against leukemia L1210 cells in culture was dramatically potentiated by the addition of guanine nucleosides to the medium. In the presence of either deoxyguanosine or guanosine, the concentration of MMPR that caused 50% inhibition of growth was 35 times lower than in the absence of these nucleosides. Similar potentiation was also observed against Sarcoma 180 cells in culture by guanosine. The metabolic basis of this synergism was approached in a study of the incorporation of [14C]glycine into 5'-phosphoribosyl-N-formylglycinamide in Sarcoma 180 cells. The results show that the site of inhibition resulting in synergism is an early step in purine biosynthesis, probably phosphoribosyl pyrophosphate amidotransferase (EC 2.4.2.14). In the L1210 cell system, the addition of hypoxanthine to the medium prevented the potentiation of MMPR by guanine nucleosides supporting the conclusion that the site of the synergistic interaction involves purine biosynthesis de novo. While hypoxanthine partially reversed the growth-inhibitory effects of MMPR, an even higher degree of protection was observed in the presence of both uridine and hypoxanthine, suggesting that MMPR may have additional sites of action concerned with pyrimidine metabolism.

Animals↗

PubMed1987

Quantitative description of nuclear morphology in assessing resistance of sarcoma 180 to adriamycin.

Resistance to adriamycin generally is explained through changes of cell/drug interactions that possibly reflect structural alterations of intracellular targets. One of the main targets of adriamycin is believed to be nuclear chromatin. In order to recognize chromatin alterations, we studied cell nuclei morphology and chromatin structure by means of digital image analysis. The studies were performed in both adriamycin-sensitive and -resistant Sarcoma 180 cell lines which were cultured under growth-stimulated and nonstimulated conditions. Using specially developed methods, we extracted parameters characterizing geometrical, optical, and structural properties of the cell nuclei from light microscopical images. The latter parameters concerned microscopical appearances of condensed chromatin and were described by features of high-optical-density regions. The results demonstrated that the quantitative criteria applied enabled the discrimination of sensitive and resistant cells. The most important parameters are the nuclear size, number, distribution, and optical density of condensed chromatin regions. In addition, the criteria permit recognition of changes related to differences in the growth conditions of the cells. The data of the image analysis suggest that adriamycin resistance in Sarcoma 180 cells is associated with characteristic patterns of cell nuclear morphology which can be described with a sufficient number of appropriate parameters. The advantages of image analysis are evident when these results are compared with the flow cytometric findings. The conclusion is that structural features of nuclear chromatin provide information essential for the assessment of drug resistance.

Animals↗

PubMed1977

Lethal and sublethal effects of hydroxyurea in relation to drug concentration and duration of drug exposure in sarcoma 180 in vitro.

The lethal and sublethal effects of hydroxyurea were studied in Sarcoma 180 in vitro in relation to drug concentration and drug exposure duration using cloning methods, radioautography, and flow microfluorometry. It was shown that postperturbation changes in radioautographic labeling intensity reflected real changes in the rate of DNA synthesis in individual cells. The data suggest that both the lethal and sublethal effects of hydroxyurea are dependent on the rate of DNA synthesis. These findings have important implications for the interpretation of DNA content distributions under perturbation conditions and for the development of drug treatment regimens that are based on cell kinetics.

Animals↗

PubMed1975

[Sensitivity tests of malignant tumours against cytostatic agents in vitro and in vivo/studies on the mouse sarcoma 180 (author's transl)].

The effects of ten different substances on the mouse sarcoma 180 have been compared using in vivo and in vitro test systems. The size of the tumours was taken as a measure of the success of the therapy in animal experiments. The in vitro effects were estimated by measuring the incorporation of radioactively labelled uridine in tumour cell suspensions from solid tumours, in ascites tumours and in tissue culture. Similar results were obtained using all three in vitro test systems. Four substances (daunomycin, fluoruracil, actinomycin D, adriamycin) exhibited activity both in vivo and in vitro, whereas five substances (cytosinarabinoside, methotrexate, ibenzmethyzin, triaziquone, bleomycin) showed no activity in any of the test systems used. With podophyllic acid ethyl hydrazide, however, no correlation between in vivo and in vitro effects was observed. Studies on the influence of the cytostatic agents on the rate of transport of uridine into the cells showed that podophyllic acid ethyl hydrazide strongly reduced the permeability of the cells to radioactive uridine.

Animals↗

PubMed1986

Effects of the antitumor agents from various natural sources on drug-metabolizing system, phagocytic activity and complement system in sarcoma 180-bearing mice.

Correlation between antitumor activity and effects on some biological properties, such as phagocytic activity of the reticuloendothelial system, the third component of complement (C3) activation, hepatic drug-metabolizing activities and pentobarbital-induced narcosis, of antitumor agents from various natural sources such as B B (Broncasma Berna), GU-P (Grifora umbellata polysaccharide), OK-432, PS-K (Polysaccharide Kureha), and RA-P (Rumex acetosa polysaccharide) were studied with female ICR mice implanted with Sarcoma 180 solid tumor. All of these agents depressed aniline hydroxylase and aminopyrine demethylase activities, prolonged the duration of pentobarbital-induced narcosis, and significantly enhanced the phagocytic activity and C3 activity. Especially, RA-P which has the strongest antitumor activity was the most effective in changing these activities. The biological activities of GU-P at a dose of 10 mg/kg reached the same level as that found with PS-K at a dose of 100 mg/kg. a possible mechanism of inhibition of Sarcoma 180 solid tumor growth by the treatment with the antitumor agents could be interpreted as due to the C3 activation, the stimulation of phagocytic activity and depression of the hepatic microsomal drug-metabolizing system in tumor-bearing mice.

Animals↗

PubMed1988

Potentiation of adriamycin cytotoxicity by dipyridamole against HeLa cells in vitro and sarcoma 180 cells in vivo.

Dipyridamole (DP) is clinically prescribed for its vasodilating and antiplatelet effects. DP also inhibits nucleoside transport and enhances cytostatic activity of antimetabolites. We obtained evidence for augmentation of the cytocidal effect of Adriamycin (ADM) by DP, both in vitro and in vivo. Nontoxic levels of DP enhanced the cytotoxicity of ADM against HeLa cells, and the 50% effective concentration of ADM was decreased 2.4-fold by DP. DP also increased the activity of ADM in clonogenic assays. Intracellular levels of ADM in the case of concomitant exposure to ADM and DP were 1.5-fold higher than in the case of exposure to ADM alone, determined using high-performance liquid chromatography. Incorporation of ADM into the cells pretreated with DP was also increased (1.4-fold), while the efflux was little affected. The growth of Sarcoma 180 tumors was prominently suppressed by the combination of ADM and DP, compared to findings with ADM alone. DP also prolonged the survival of Sarcoma 180 tumor-bearing mice, when given in combination with ADM. While the enhancement of cytostatic activity of antimetabolites has been attributed to a decrease in utilization of the salvage pathway by DP, our data show that the synergic effects of DP with ADM were the result of increased intracellular levels of ADM.

Animals↗