Ehrlich Ascites Carcinoma In Mice Research Articles

Pharmacological study on Panax ginseng C. A. Meyer. XV. Effects of 70% methanolic extract from red and white ginseng on the antitumor activity of mitomycin C

The influence of various fractions and ginsenosides from the 70% methanolic extract (RMe) of Red Ginseng (a steamed and dried root of Panax ginseng C. A. Meyer) on the cytocidal effect of mitomycin C (MMC) against Ehrlich ascites carcinoma was investigated in vitro. The AcOEt soluble portion (RMe-I) showed an increasing effect on the activities of lysosomal enzymes in the cultured tumor cells. RMe-I promoted the uptake of MMC into the tumor cells and enhanced the cytotoxicity of MMC against the cultured tumor cells. 20(S)-, 20(R)-ginsenoside Rg3 and ginsenoside Rh2 isolated from RMe-I promoted the uptake of MMC into the tumor cells but ginsenosides from the n-BuOH soluble portion (RME-II) had no effect. Furthermore, the influence of RMe and the 70% methanolic extract (WMe) from White Ginseng (a dried root of Panax ginseng C. A. Meyer) on the cytocidal effect of MMC was investigated in vivo. MMC combined with RMe showed stronger antitumor effects against the ascites form of mouse Ehrlich ascites carcinoma and rat ascites hepatoma AH 130 than MMC combined with WMe. The activities of lysosomal enzymes in tumor cells were also more increased in comparison with that combined MMC and WMe.

Isolation and characterization of an antitumor active agar-type polysaccharide of Gracilaria dominguensis

Cold water extraction of the red alga Gracilaria dominguensis, followed by cetyltrimethylammonium bromide fractionation, gave a highly sulfated, agar-type polysaccharide which inhibited the transplantation of Ehrlich ascites carcinoma in mice. The structure of the polysaccharide has been investigated by methylation analysis, and 1H- and 13C-n.m.r. spectroscopy, and was shown to be mainly composed of alternating (1→3)-linked β- d-galactopyranosyl 6-sulfate and (1→4)-linked 3,6-anhydro-α- l-galactopyranosyl residues.

Terpenoids. LIII. Antitumor activity of trichorabdals and related compounds.

Among the three types, enmein-, oridonin-, and trichorabdal-type, of diterpenoids from Rabdosia trichocarpa, the latter showed the highest antitumor activity against Ehrlich ascites carcinoma in mice. Their potent activities were attributed to synergistic increase arising from the presence of two active sites in one molecule. In vitro activity against HeLa cells and in vivo activity against P 388 lymphocytic leukemia of diterpenoids and related compounds were also determined, but no synergistic increase in activity due to plural active sites was observed in those cases.

Potentiation of the antitumor activity of cisplatin in mice by 3-aminobenzamide and nicotinamide.

3-Aminobenzamide (3AB) and nicotinamide (NA), inhibitors of adenosine-ribose transferase (ADPRT), potentiated the antitumor activity of cisplatin (DDP) on Ehrlich ascites carcinoma in mice. The mean survival times of the mice increased from 21.2-37.0 days in DDP-treated groups to 47.0-54.6 days in mice treated with DDP plus NA or 3AB. These drugs also potentiated DDP antitumor activity on sarcoma 180, with the inhibition rates increasing from 12.4%-20.8% in groups treated daily with DDP to 29.8%-46.4% in those treated with DDP plus NA or 3AB; however, neither 3AB nor NA alone showed any antitumor activity. The single-dose lethality of DDP on mice was partially reversed by either NA or 3AB. The pathological study revealed that the morphologic changes in the proximal tubules 1 month after a single dose of DDP (10 mg/kg) were partially prevented by a single protective dose (5 mmol/kg) of NA or 3AB. Our results suggest that the combination of DDP with ADPRT inhibitors might be used clinically in the future.

The Antitumor Effect of Bleomycin Combined with Bestatin against Ehrlich Ascites Carcinoma in Mice

The effect of bleomycin against Ehrlich ascites carcinoma transplanted subcutaneously to mice used in combination with bestatin was investigated. Male Balb/c mice weighting approximately 20 g and bred in our laboratories were used in this study. Each mouse was injected in its left lateral abdominal region subcutaneously with 7 X 10(6) tumor cells in 0.2 ml of ascites fluid. The mice were divided into four groups: control, bestatin alone (5 mg/kg intraperitoneally on Days 9-14), bleomycin alone (10 mg/kg intraperitoneally on Days 7 and 8), and bestatin plus bleomycin. Our results show that bestatin enhances the antitumor effect of bleomycin against Ehrlich ascites carcinoma as measured by the increased survival rates. Being an agent of very low toxicity, bestatin should be considered as a part of the chemoimmunotherapy protocol.

The purification and preliminary characterization of UMP synthase from human placenta.

Orotate phosphoribosyltransferase (EC 2.4.2.10) and orotidine 5'-monophosphate decarboxylase (EC 4.1.1.23) are the final two of six enzymatic steps required in the de novo biosynthesis of uridine 5'-monophosphate (UMP). Earlier work of this laboratory showed that, in the mouse Ehrlich ascites carcinoma, both of these enzymatic activities were contained on the single multifunctional polypeptide chain, UMP synthase. We report here that the placenta provided an available human source for UMP synthase with 40-fold higher orotate phosphoribosyltransferase and orotidine 5'-monophosphate decarboxylase specific activities than erythrocytes, a human source previously used by others. By using the placenta as a source of UMP synthase and by developing a novel purification procedure different from that used in the purification of UMP synthase from the Ehrlich ascites carcinoma (the only other homogeneous preparation of a mammalian UMP synthase), we achieved the purification of human UMP synthase to apparent homogeneity. This represents the first publication to homogeneity of UMP synthase from a human source as well as from a source other than malignant cell lines. Contrary to earlier reports human placental UMP synthase was found to be a multifunctional protein containing both enzymatic activities on a single polypeptide of 51,000 molecular weight. Preliminary characterization of the human placental UMP synthase showed it to be similar to UMP synthase from the Ehrlich ascites carcinoma in subunit molecular weight, native molecular weight, isozyme pattern (although not absolute pI values), pH optima of enzymatic activities, and kinetic constants for orotidine 5'-monophosphate (Km) and 6-azauridine 5'-monophosphate (Ki) at the decarboxylase site.

Coordination metal complexes of Rh(I), Ir(I) and Ru(II): recent advances on antimetastatic activity on solid mouse tumors

Following pioneering observations on the mouse Ehrlich ascites carcinoma, the result of cooperation between the Institutes of Chemistry and Pharmacology of the University of Trieste, more recent studies have characterized some aspects of the antimetastatic properties of coordination metal complexes other than platinum compounds. The compounds examined, rhodium(I) and iridium(I) derivatives of the type [Mchel(LL)] +/o (chel= pyridinalimine (NNR), acetylacetonate; LL= 1,5-hexadiene, 1,5-cyclooctadiene, norbornadiene), both with a square-planar structure, and an octahedral ruthenium(II) derivative RuCl 2(dimethylsulfoxide) 4 were tested using the solid metastasizing tumor of the mouse, Lewis lung carcinoma. The conclusions which can be drawn from the resulting data concern the role of the metal, of the leaving group and of the non-leaving group as well. It was found that the organometallic complexes of Rh(I) are more active than those of Ir(I); within the Rh(I) derivatives of the type [Rh(I)COD(NNR)] +Cl − (RCH 3, C 2H 5, iC 3H 7), the higher the hydrosolubility, the higher is the antitumor and particularly the antimetastatic effect; as far as the diolefinic ligands are concerned, the higher the chelating effect the higher are the antimetastatic properties of the resulting compound. Separate conclusions can be made for the Ru(II) derivative. Comparison of its antimetastatic effects with those of cis-dichlorodiammineplatinum(II) (cisplatin) using three solid mouse tumors, clearly shows a better therapeutic index for the former, suggesting that within this class of compounds it is conceivable to obtain derivatives with an antineoplastic activity comparable to or even higher than that of cisplatin.

Effect of several d-morphinans on ascites tumors in mice.

Dextromethorphan and its analogues (DM 16, DM 34, DM 72, DM 75 and DM 96) were examined for their effect on Ehrlich ascites carcinoma or ascites sarcoma-180 in female mice of the ddY strain. The suspension of Ehrlich carcinoma cells or sarcoma-180 cells was prepared from mice at 10 days after i.p. inoculation of the cells, using Hanks' balanced salt solution, and the cell suspension was inoculated i.p. into mice (2 X 10(6) viable cells/mouse). The chemicals dissolved in physiological saline containing 5% HCO-60 were then injected i.p. into the mice once daily for 5 successive days (5-40 mg/kg/day). In addition, mice given the tumor cells were treated with the saline containing 5% HCO-60 alone for 5 days (untreated mice). In groups of mice bearing Ehrlich ascites carcinoma or ascites sarcoma-180, the mean survival time of mice treated with 20-40 mg/kg/day of DM 96 was more than twice that of the corresponding untreated mice. The mean survival time of mice treated with 20 mg/kg/day of DM 96 was also longer than that of mice treated with 40 mg/kg/day of the other chemicals, irrespective of the ascites tumors. Concerning these survival times, the LD50 (i.p.) of DM 96 in mice differed slightly from that of other chemicals (88 mg/kg and 77-106 mg/kg). These results indicate that DM 96 is more active than the other chemicals against the ascites tumors in mice.

Antitumor effect of fibrinogen microparticles containing adriamycin on Ehrlich ascites carcinoma in mice.

The antitumor activity of fibrinogen microparticles containing adriamycin against Ehrlich ascites carcinoma in mice was evaluated on the basis of animal survival data. Tumor cell injections were performed on day 0 and microparticle injections on day 1, both intraperitoneally. A prolongation of the life span of tumor-bearing mice following injection of therapeutic microparticles was noted, and the microparticles containing adriamycin were therapeutically more effective than adriamycin alone. The high chemotherapeutic efficiency of fibrinogen-adriamycin microparticles was striking at high doses which would be toxic in the case of the drug alone. These results indicated that fibrinogen microparticles containing adriamycin may be effective in cancer chemotherapy. Microparticles composed of fibrinogen could be a novel drug carrier for use in injectable delivery systems for anticancer agents.

Preparation and evaluation in vitro and in vivo of fibrinogen microspheres containing adriamycin.

Fibrinogen microspheres containing adriamycin were prepared and eveluated as a novel drug delivery system. The shape of the microspheres was invariably spherical. The average diameters were 2.4 and 1.7 μm, and the drug contents in the microspheres were 10.2 and 9.9%, for the microspheres prepared at 110 and 140°C, respectively. Release of the drug from fibrinogen microspheres was slow, and the drug release continued over. 7 d. These results indicated that drug-loaded fibrinogen microspheres could provide a sustained release property in vitro.The antitumor activity of fibrinogen microspheres containing adriamycin was evaluated against Ehrlich ascites carcinoma in mice on the basis of animal survival data. Tumor cell injections were performed on day 0 and microsphere injections on day 1, both intraperitoneally. A prolongation of the life span of tumor-bearing mice following injection of therapeutic microspheres was noted, and the microspheres containing adriamycin were therapeutically more active than adriamycin alone. The high chemotherapeutic efficiency of fibrinogen-adriamycin microspheres was striking at high doses, which would be toxic in the case of the drug alone.These results indicated that fibrinogen microspheres containing adriamycin may be effective in cancer chemotherapy. Microspheres composed of fibrinogen show good biocompatibility and could be useful as a novel drug carrier in injectable delivery systems for anticancer agents.

Fibrinogen microspheres as novel drug delivery systems for antitumor drugs.

Fibrinogen microspheres containing 5-fluorouracil (5-FU) were prepared and evaluated as a novel drug delivery system. The shape of the microspheres was invariably spherical. The average diameters were 3.9 and 1.1 μm and the drug contents in the microspheres were 16.1 and 6.1%, for the microspheres prepared at 90 and 160°C, respectively. Release of the drug from fibrinogen microspheres was slow after the initial burst, and the drug release continued over 5d. These results indicated that the drug-loaded fibrinogen microspheres could be prepared providing a sustained release property in vitro. The antitumor activity of fibrinogen microspheres containing 5-FU was evaluated against Ehrlich ascites carcinoma in mice. The mean survival time of the group receiving the microspheres was larger than that of the control group, though essentially the same as that of the free 5-FU-treated group. Fibrinogen microspheres show good biocompatibility and should be useful as a novel drug carrier in injectable delivery systems for antitumor drugs.

Purification and properties of mouse thymus thymidylate synthase. comparison of the enzyme from mammalian normal and tumour tissues

1. 1. Mouse thymus thymidylate synthase has been purified to apparent electrophoretic homogeneity and compared with the enzyme from mouse tumour L1210 and Ehrlich ascites carcinoma cells. 2. 2. The enzyme is a dimer composed of 35,000 mol. wt monomers. 3. 3. Mouse thymus and tumour enzymes exhibit allosteric properties reflected by cooperative binding of both dUMP and 5-fluoro-dUMP. 4. 4. Activation energy for the reaction, catalyzed by thymidylate synthase from mouse tumour but not from mouse thymus, lowers at temperatures above 34† C, reflecting a change of rate-limiting step in dTMP formation. 5. 5. MgATP at millimolar concentrations inhibits mouse thymus enzyme.

Antitumor effect of implanted ethylene-vinyl alcohol copolymer matrices containing anticancer agents on Ehrlich ascites carcinoma and P388 leukemia in mice.

Ethylene-vinyl alcohol (EVAI) copolymer was evaluated as a vehicle for controlled or sustained release of 5-fluorouracil (5-FU) and adriamycin. The variation of initial drug concentration and, to an even greater extent, the variation in comonomer ratio affected the drug release rate as well as the cumulative amount of the drug released. An increase in the ethylene content of the EVAI copolymer decreased drug release. It appears that EVAI copolymer can be used as a membrane for controlling the release of 5-FU or adriamycin. The antitumor activity of EVAI copolymer matrices containing these anticancer agents was evaluated against Ehrlich ascites carcinoma (EAC) in mice on the basis of changes in body weight and animal survival data. Tumor cell injections were performed on day 0 and matrix implantations on day 3, both intraperitoneally. The suppressive effect of matrices containing anticancer agents on the increase in body weight was higher than that of the free drugs. A prolongation of the life span of tumor-bearing mice following implantation of therapeutic matrices was also noted. Implantation of the EVAI copolymer matrix containing an anticancer agent was less effective against the P388 leukemia than the EAC. These results suggest that EVAI copolymer matrices containing anticancer agents may be effective in cancer chemotherapy. Matrices composed of EVAI copolymer could be useful vehicles for implanted delivery systems for anticancer agents.

Antitumor activity of 7-(hydroxyamino)-5,8-quinolinedione

7-(Hydroxyamino)-5, 8-quinolinedione (7-HAQD), a simple mimetic compound of Streptonigrin in whose chemical structure an aminoquinone moiety has been recognized to be significant for the appearance of its antitumor activity, was investigated for antitumor activities. 7-HAQD showed difference spectra toward deoxyribonucleic acid (DNA), and inhibited their precursor incorporation into DNA and ribonucleic acid (RNA) in Ehrlich ascites carcinoma cells. Also, 7-HAQD inhibited against the proliferation of L1210 leukemia cells in vitro. 7-HAQD exhibited distinct antineoplastic effects against Ehrlich mouse ascites carcinoma and P388 leukemia. However, its inhibitory effects against the solid carcinomas of Ehrlich and Sarcoma 180 were weaker than those against the ascites carcinomas.

Stimulation of RNA polymerases I and II from mouse L1210 leukemia and Ehrlich ascites carcinoma cells by spermine and spermidine and its modification by ammonium sulfate

1. 1. Nuclear RNA polymerases from murine L1210 leukemia and Ehrlich carcinoma cells were stimulated more effectively by spermine than by spermidine. 2. 2. Optimal stimulatory concentrations of spermine and spermicline for Ehrlich polymerases Ia and Ib decreased to physiological values and maximal stimulation increased as the concentration of (NH 4) 2SO 4 was reduced from 0.08 to 0 M. 3. 3. In the presence of 0.062–0.074M (NH 4) 2SO 4 L1210 polymerases Ia, IIa and IIb were stimulated significantly by both polyamines, whereas, at (NH 4) 2SO 4 concentrations of 0.11–0.17 M, stimulation was suppressed and high concentrations of the polyamines were inhibitory. 4. 4. Similarly, stimulation of Ehrlich solubilized polymerase by polyamines was inhibited by 0.064 M (NH 4) 2SO 4.

Induction by 5-fluorouracil of a major phase difference in the circadian profiles of DNA synthesis between the Ehrlich ascites carcinoma and five normal organs

DNA synthetic activity (DNA-SA) was measured by the incorporation of tritiated thymidine (TDR) into chemically isolated DNA. DNA-SA was monitored every 3 h over a 60-h period in the Ehrlich ascites carcinoma (EAC), tongue, bone marrow, ileum, stomach and rectum in mice which had received 50 mg/kg 5-fluorouracil (FU) at 17:00 h. FU was given in an attempt to induce major phase differences in the circadian profiles of DNA-SA between the EAC and the normal organs. Subsequent to the FU treatment, a time period 15 h in duration, was identified when DNA-SA in the EAC was at or near a maximal level (125–190 cpm/μg DNA) whereas DNA-SA in all 5 normal organs was at or near a minimal level (< 60 cpm/μg DNA). Induction of such quantitative (actual levels of DNA-SA) and qualitative (180° circadian phase change) differences between the normal and neoplastic cells in the same host should be useful in designing a more effective chemotherapeutic protocol for this tumor-host situation.

Pimaricin potentiation of bleomycin activity against murine tumors.

Antitumor activity of bleomycin against Ehrlich ascites carcinoma in mice was examined in the presence and absence of a polyene antibiotic, pimaricin. A single intraperitoneal (ip) injection of bleomycin and pimaricin in combination into tumor-bearing mice on day 1 produced a synergistic life-prolongation effect as compared with that of each agent alone. Accumulation of abdominal ascites was almost completely blocked by bleomycin in combination with the tetraene antibiotic. Frequent ip injection of small doses of bleomycin and pimaricin on day 1 produced a much greater increase in life span than the single ip injection. To obtain effective synergism under these treatment schedules, it appeared that the dose of bleomycin should be reciprocally proportional to the dose of pimaricin.

Studies on antitumor activity of Bas idiomycete polysaccharide (Report 32) : Antitumor effect and the mechanism of ATOM prepared from cultured Agari cus blazei

An antitumor polysacchari de, a mannan preparation (ATOM : antitumor organic substance Mie) isolated from cultured Agaricusblazei, suppressed markedly the growth of solid Sarcoma-180 and Ehrlich ascites carcinoma in mice. In combination therapy with mitomycin C, 5-f1uorouraci1 and cytosine arabinoside, ATOM showed a remarkable synergistic effect against these tumors. ATOM alone did not show direct cytocidal activity against HeLa-S3, Ehrlich ascites carcinoma and Sarcoma-180 ascites cells i_n_ vitro. ATOM showed strongly enhanced phagocytosis of Salmone!1 a enteri ti di s No.11 by peritoneal macrophages of mouse after i.p. injection.

Effect of liposomal encapsulation of cis-platinum diamminodichloride in the treatment of Ehrlich ascites carcinoma.

cis-Platinum diamminodichloride (II) encapsulated in neutral liposomes has been used to enhance the activity of the drug on Ehrlich ascites carcinoma in mice. Both in vitro and in vivo experiments have revealed that better regression in tumor growth and cure, could be achieved with a lower dose, having apparently no toxic effect, by liposomal encapsulation of the drug.

Potential for ambiguity in the interpretation of immunochemical studies on multienzymic proteins: The case of uridylate synthase

Rabbit antibodies directed against homogeneous uridylate synthase multienzyme from mouse Ehrlich ascites carcinoma precipitate both the orotidine-5′-monophosphate decarboxylase (EC 4.1.1.23) and orotate phosphoribosyltransferase (EC 2.4.2.10) activities of mouse and human erythrocyte uridylate synthase. When the partially purified human enzyme is used as antigen the two activities coprecipitate with the same apparent titer; however, when the mouse carcinoma protein was studied under the same conditions the decarboxylase activity immunoprecipitated with significantly higher avidity than did the transferase activity. Since the mouse multienzyme has been shown to be a single polypeptide that contains both activities (McClard, R.W., Black, M.J., Livingstone, L.R. and Jones, M.E. (1980) Biochemistry 19, 4699–4706), these results were, at face value, surprising. However, when the mouse orotate phosphoribosyltransferase activity (which is largely lost upon dilution into the immunoassay medium) was stabilized with 5-phosphoribosyl 1-pyrophosphate, both enzyme activities displayed the same apparent antibody titer. The immunochemical studies indicate that the antibodies, as a population, preferentially bind to a form or forms of the enzyme which contain(s) denatured transferase domains. A calculation based on a simple model yields a value of approximately 100 for the relative selectivity of the antibody for the denatured form of uridylate synthase. These results illustrate an ambiguity that is inherent in the interpretation of immunochemical studies on such multienzymic proteins; that is, it is possible to conclude incorrectly that two enzyme activities are not functionally associated if one of the catalytic domains is particularly unstable and thereby displays greater immunoreactivity for the specific antiserum.