Mouse Ascites Tumor Research Articles

Carbo animalis and immune response to Ehrlich ascites tumor in mice: an experimental model

The quality of life in cancer patients is largely related to the activity of the immune system. Clinical reports show improvement in the quality of life of terminally ill patients treated with the homeopathic medicine carbo animalis, however, the literature on this subject is rare. The objective of this study was to propose an experimental model to study the possible effects of carbo animalis in the immune response to a highly malignant carcinoma, as well as their impact on the general condition of the sick animals. Male Balb/c mice were inoculated with Ehrlich ascites tumor and treated with carbo animalis 6cH or 6cH + 30cH (potency association). The control group was treated with the same succussioned vehicle. Clinical signs, survival and the local immune response (peritoneal) were evaluated. T lymphocytes, B1 and B2, NK cells and phagocytes were identified and quantified by immuno-cytochemistry and flow cytometry. Animals treated with carbo animalis 6cH + 30cH showed increase of incidence in clinical signs comparing to the other groups. The local immune response, showed increase in the proportion of CD25+ cells in relation to total T cells and increase of B1 cells compared to B2 cells in the group treated with carbo animalis 6cH. In contrast, animals treated with carbo animalis 6cH + 30cH showed increase in the number of CD3+ cells and NK cells, both adhered to tumor cells. Although the clinical significance of these findings are still under discussion, this preliminary work provides a useful experimental protocol for the study of the mechanisms of this remedy and shows the possible relevance of homeopathic potencies association in the anti-neoplasm treatments.

Chronic Dietary Administration of the Glycolytic Inhibitor 2-Deoxy-D-Glucose (2-DG) Inhibits the Growth of Implanted Ehrlich’s Ascites Tumor in Mice

BackgroundDietary energy restriction (DER) has been well established as a potent anticancer strategy. Non-adoption of restricted diet for an extended period has limited its practical implementation in humans with a compelling need to develop agents that mimic effects similar to DER, without reduction in actual dietary intake. Glycolytic inhibitor, 2-deoxy-D-glucose (2-DG), has recently been shown to possess potential as an energy restriction mimetic agent (ERMA). In the present study we evaluated the effect of dietary 2-DG administration on a mouse tumor model, with a focus on several potential mechanisms that may account for the inhibition of tumorigenesis.Methodology/Principal FindingsSwiss albino strain ‘A’ mice were administered with 0.2% and 0.4% w/v 2-DG in drinking water for 3 months prior to tumor implantation (Ehrlich’s ascites carcinoma; EAC) and continued till the termination of the study with no adverse effects on general physiology and animal growth. Dietary 2-DG significantly reduced the tumor incidence, delayed the onset, and compromised the tumor growth along with enhanced survival. We observed reduced blood glucose and serum insulin levels along with decreased proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine positive (BrdU+) tumor cells in 2-DG fed mice. Also, reduced levels of certain key players of metabolic pathways such as phosphatidylinositol 3-kinase (PI3K), phosphorylated-Akt and hypoxia inducible factor-1 alpha (HIF-1α) were also noted in tumors of 2-DG fed mice. Further, decrease in CD4+/CD8+ ratio and T-regulatory cells observed in 2-DG fed mice suggested enhanced antitumor immunity and T cell effector function.Conclusion/SignificanceThese results strongly suggest that dietary 2-DG administration in mice, at doses easily achievable in humans, suitably modulates several pleotrophic factors mimicking DER and inhibits tumorigenesis, emphasizing the use of ERMAs as a promising cancer preventive strategy.

Antitumor activity of two Streptomyces extracts (Ag18 & Ag20) on Ehrlich ascites tumor in mice in vitro and in vivo studies

Antitumor activity of two Streptomyces extracts (Ag18 & Ag20) on Ehrlich ascites tumor in mice in vitro and in vivo studies

Modulation of Cytokines Production by Indomethacin Acute Dose during the Evolution of Ehrlich Ascites Tumor in Mice.

The aim of the present study was to investigate the influence of a nonselective COX1/COX2 inhibitor (indomethacin) on tumor growth of Ehrlich Ascites Tumor (EAT) in mice, using as parameters the tumor growth and cytokine profile. Mice were inoculated with EAT cells and treated with indomethacin. After 1, 3, 6, 10, and 13 days the animals were evaluated for the secretion of TNFα, IL-1α, IL-2, IL-4, IL-6, IL-10, and IL-13 and PGE2 level in peritoneal cavity. The results have shown that EAT induces PGE2 production and increases tumor cells number from the 10th day. The cytokine profile showed EAT induces production of IL-6 from 10th day and of IL-2 on 13th day; the other studied cytokines were not affected in a significant way. The indomethacin treatment of EAT-bearing mice inhibited the tumor growth and PGE2 synthesis from the 10th day. In addition, the treatment of EAT-bearing mice with indomethacin has stimulated the IL-13 production and has significantly inhibited IL-6 in the 13th day of tumor growth. Taken together, the results have demonstrated that EAT growth is modulated by PGE2 and the inhibition of the tumor growth could be partly related to suppression of IL-6 and induction of IL-13.

Dual effects of indoleamine 2,3-dioxygenase inhibitors on the therapeutic effects of cyclophosphamide and cycloplatam on Ehrlich ascites tumor in mice.

Ethyl pyruvate, an inhibitor of indoleamine 2,3-dioxygenase, slightly suppressed the growth of transplantable Ehrlich tumor in mice and significantly potentiated the therapeutic effect of cyclophosphamide. Another inhibitor amidoxime produced a similar effect. However, both ethyl pyruvate and amidoxime significantly reduced the effect of cycloplatam therapy. The observed changes can be stipulated by different effects of cyclophosphamide and cycloplatam on the subpopulations of lymphoid cells taking part in the formation of antitumor immunity and resistance to tumors.

Substituted 3‑acyl‑2‑phenylamino‑1,4‑naphthoquinonesintercalate into DNA and cause genotoxicity through the increasedgeneration of reactive oxygen species culminating in cell death.

Naphthoquinones interact with biological systems by generating reactive oxygen species (ROS) that can damage cancer cells. The cytotoxicity and the antitumor activity of 3‑acyl‑2‑phenylamino‑1,4‑naphthoquinones (DPB1‑DPB9) were evaluated in the MCF7 human breast cancer cell line and in male Ehrlich tumor‑bearing Balb/c mice. DPB4 was the most cytotoxic derivative against MCF7 cells (EC50 15µM) and DPB6 was the least cytotoxic one (EC50 56µM). The 1,4‑naphthoquinone derivatives were able to cause DNA damage and promote DNA fragmentation as shown by the plasmid DNA cleavage assay (FII form). In addition, 1,4‑naphthoquinone derivatives possibly interacted with DNA as intercalating agents, which was demonstrated by the changes caused in the fluorescence of the DNA‑ethidium bromide complexes. Cell death of MCF7 cells induced by 3‑acyl‑2‑phenylamino‑1,4‑naphthoquinones was mostly due to apoptosis. The DNA fragmentation and subsequent apoptosis may be correlated to the redox potential of the 1,4‑naphthoquinone derivatives that, once present in the cell nucleus, led to the increased generation of ROS. Finally, certain 1,4‑naphthoquinone derivatives and particularly DPB4 significantly inhibited the growth of Ehrlich ascites tumors in mice (73%).

Synergism Between Propolis and Hyperthermal Intraperitoneal Chemotherapy with Cisplatin on Ehrlich Ascites Tumor in Mice

We investigated antitumor, genotoxic, chemopreventive, and immunostimulative effects of local chemoimmunotherapy and hyperthermal intraperitoneal chemotherapy (HIPEC) in a mouse-bearing Ehrlich ascites tumor (EAT). Mice were treated with water-soluble derivative of propolis (WSDP) at a dose of 50 mg kg(-1) , 7 and 3 days before implantation of EAT cells, whereas cisplatin (5 or 10 mg kg(-1) ) was injected 3 days after implantation of EAT cells at 37°C and 43°C. The following variables were analyzed: the total number of cells, differential count of the cells present in the peritoneal cavity, functional activity of macrophages, comet assay, and micronucleus assay. The combination of WSDP + CIS 5 mg kg(-1) at 37°C resulted in tumor growth inhibition and increased the survival of mice by additional 115.25%. WSDP with HIPEC increased the survival of mice by additional 160.3% as compared with HIPEC. WSDP reduced cisplatin toxic and genotoxic effect to normal cells without affecting cisplatin cytotoxicity on EAT cells. In addition, WSDP with HIPEC increased the cytotoxic actions of macrophages to tumor cells. Water-soluble derivative of propolis increases macrophage activity and sensitivity of tumor cells to HIPEC and reduces cisplatin toxicity to normal cells.

Transdermal delivery of adriamycin to transplanted ehrlich ascites tumor in mice.

There is considerable interest in the skin as a site of anti-cancer drug application. Nevertheless, the skin poses a formidable barrier to drug penetration, thereby limiting topical and transdermal bioavailability. However, we previously showed that a thioglycolate-based depilatory agent increases the drug permeability of mouse skin. In the present report, we investigated the skin permeability and efficacy of the anti-cancer drug adriamycin increased when administered transdermally to mice in combination with a thioglycolate-based depilatory agent. Adriamycin in combination with depilatory treatment reduced Ehrlich tumor growth in hairless mice about the weight and size of harvested tumors. In addition, our delivery method for adriamycin increased the therapeutic effectiveness of this agent by decreasing toxicity. Moreover, measurement of adriamycin autofluorescence revealed that topically applied adriamycin penetrate the dermis after depilatory agent treatment. These results indicate that the transdermal delivery of anti-cancer drugs is feasible by handy pretreatment of the skin with a thioglycolate-based depilatory agent.

The inhibitory effect of <i>Binens bipinnata L</i>. extract on U14 tumour in mice

The objective of this paper was to study the in vitro and in vivo inhibitory effect of Bidens bipinnata L. extract on growth of cervical carcinoma U14 cells. MTT method was used to determine the inhibitory effect of Bidens bipinnata L. extract on U14 tumour cells, and the effects of Bidens bipinnata L. extract on inhibition rate of solid tumour and life prolongation rate of ascites tumour were observed through the establishment of two animal models of mouse cervical carcinoma U14 solid tumour and ascites tumour. In the in vitro MTT assay, the inhibition rate gradually increased with the increase of dose of Bidens bipinnata L. and the extension of time. Its inhibition rate was 70.44% at a concentration of 80µg/L. Solid tumour inhibition rates in the high- and low-dose groups and cisplatin group were 49.13%, 2.26% and 75.72% respectively; life prolongation rates in each ascites tumour group were 63.63%, 34.86% and 87.34% respectively. The Bidens bipinnata L. extract has a certain inhibitory effect on growth of mouse cervical carcinoma U14.

Preliminary Evaluation of In Vitro Cytotoxicity and In Vivo Antitumor Activity of Xanthium strumarium in Transplantable Tumors in Mice

In the present study, active fractions of the methanolic extract of Xanthium strumarium (XS) showing potent cytotoxicity were determined using microculture tetrazolium (MTT) and sulforhodamine B (SRB) assays in selected cancer cell lines. The active fractions viz., chloroform soluble fraction of root (CEXSR), hexane soluble fraction of leaf (HEXSL), hexane soluble fraction of fruits (HEXSF) and chloroform soluble fraction of fruits (CEXSF) of XS were tested in transplantable animal tumor models for their antitumor potential. Dalton's ascitic lymphoma (DLA) cells were used to induce solid and liquid (ascites) tumor in mice. The tumor bearing animals were treated with active fractions at two dose levels (100 and 200 mg/kg). The antitumor activities of the active fractions in tumor bearing animals were monitored with parameters such as body weight and increase in life-span as well as biochemical and hematological modalities (in the case of liquid tumor). Tumor incidence and tumor volume were the parameters monitored in the case of the solid tumor model. The results were analyzed by one-way ANOVA followed by Tukey's post hoc test. The extracts were found to increase the life-span of tumor bearing animals and restore the altered hematological and biochemical parameters significantly.

Enhancement of Antitumor Effects of a New Boron Compound Combined with Ultrasound on the Mouse Ascites Tumor

Enhancement of Antitumor Effects of a New Boron Compound Combined with Ultrasound on the Mouse Ascites Tumor

Enhancement of Antitumor Effects of a New Boron Compound Combined with Ultrasound on the Mouse Ascites Tumor

The use of ultrasound in therapeutic medicine is a developing field. The action of many anticancer agents can be improved with the use of ultrasonic exposure either in vitro or in vivo. Boron-based compounds have shown promising results as chemotherapeutic agents in specific forms of cancer. We synthesized a new boron compound of chloroacetohydroxamic acid, 2-hydroxyamino-oxoethylborate (HAOB). HAOB was used in combination with ultrasound (US) at a frequency of 25 KHz, and inhibitory effects on the growth of mouse ascites tumours were monitored by observing survival time, tumour growth, cell viability and DNA synthesis of treated Ehrlich ascites carcinoma (EAC) cells. 5-Fluorouracil (5-FU) was used as a positive control. The results indicated that HAOB alone produced significant antitumor action whereas combination of US with HAOB or 5-FU produced an extra antitumor action when compared to the actions of HAOB or 5-FU used alone.

Acceleration of Anaerobic Cysteine Transformations to Sulfane Sulfur Consequent toγ-Glutamyl Transpeptidase Inhibition

Toxicity of drugs and radiation in the cells is largely dependent on the level of thiols. In the present studies, an attempt has been made to inhibit γ-glutamyl transpeptidase (γGT) activity in EAT-bearing animals tissue. We have expected that administration of γGT inhibitors: acivicin and 1,2,3,4-tetrahydroisoquinoline (TIQ) may influence GSH/γ–glutamyl transpeptidase (γGT) system in the regulation of cysteine concentration and anaerobic cysteine metabolism in normal and cancer cells. Development of Ehrlich ascites tumor in mice enhances peroxidative processes, diminishes levels of nonprotein thiols (NPSH) and sulfane sulfur, and lowers activities of enzymes involved in its formation and transfer in the liver and kidney. Although γGT inhibitors further decrease NPSH level, they increase cysteine and sulfane sulfur levels. This means that upon γGT inhibition, cysteine can be efficiently acquired by normal liver and kidney cells via another pathway, that is so productive that sulfane sulfur level and intensity of anaerobic cysteine metabolism even rise.

Use of Curcumin to Decrease Nitric Oxide Production During the Induction of Antitumor Responses by IL-2

Nitric oxide (NO) synthesis is strongly induced during interleukin (IL)-2 treatment of mice and humans. Although this free radical can act as a cytotoxic effector molecule against cancer cells, immunosuppressive effects have also been suggested. We evaluated the effects of curcumin on IL-2-induced NO synthesis and IL-2-induced antitumor responses in a mouse ascites tumor model. Curcumin inhibited inducible nitric oxide synthase (iNOS) expression and NO production, and thereby enhanced the proliferation and cytotoxic activity of cocultured lymphocytes and macrophages during IL-2 stimulation which we earlier established as an in vitro model of IL-2-induced NO synthesis. Curcumin also decreased apoptosis of cocultured lymphocytes and macrophages during IL-2 stimulation. In contrast, the curcumin-induced changes in proliferation and apoptosis were not observed in cultures of lymphocytes alone, macrophages alone, and cocultured lymphocytes/iNOS-knock out macrophages, all of which produced little nitrite during IL-2 stimulation. In conjunction with IL-2 treatment, oral curcumin administration significantly inhibited IL-2 therapy-induced urinary nitrite/nitrate excretion and iNOS expression of tumor tissues, and further increased the IL-2 therapy-induced prolongation of survival in a murine Meth-A ascites tumor model. Curcumin may be useful as an adjunct to increase the antitumor activity of IL-2 therapy.

Improved therapeutic activity of folate-targeted liposomal doxorubicin in folate receptor-expressing tumor models

The folate receptor (FR) is overexpressed in a broad spectrum of malignant tumors and represents an attractive target for selective delivery of anti-cancer agents to FR-expressing tumors. Targeting liposomes to the FR has been proposed as a way to enhance the effects of liposome-based chemotherapy. Folate-polyethylene glycol-distearoyl-phosphatidyl-ethanolamine conjugate was inserted into pegylated liposomal doxorubicin (PLD). The therapeutic activity of folate-targeted (FT-PLD) and non-targeted (PLD) pegylated liposomal doxorubicin was tested in two human tumor models (KB, KB-V) and in one mouse ascitic tumor model (FR-expressing J6456) by the i.v. systemic route in all models, and by the i.p. intracavitary route in the ascitic tumor model only. Consistent with previous studies, PLD was clearly superior to free doxorubicin in all tumor models. When targeted and non-targeted liposome formulations were compared, FT-PLD was more effective than PLD in the KB and KB-V xenograft models, and in the J6456 intra-cavitary therapy model. The therapeutic effect was dose-dependent in the KB model and schedule-dependent in the J6456 intra-cavitary therapy model. In some experiments, toxic deaths aggravated by folate-depleted diet were a major confounding factor. In a non-FR expressing J6456 model, FT-PLD was as active as PLD indicating that its activity is not limited to FR-expressing tumors. Folate-targeting confers a significant albeit modest therapeutic improvement to PLD in FR-expressing tumor models, which appears particularly valuable in intracavitary therapy. The potential clinical added value of this approach has yet to be determined.

Chromosomal evolution in the Ehrlich-Lettré complex of hyperdiploid mouse ascites tumors: results from seven laboratory strains.

The chromosomes of 7 strains of the hyperdiploid Ehrlich-Lettre mouse ascites tumor (ELD) were analyzed by conventional stainings and by G- and C-bandings. A tendency to slow decrease in chromosome number during a 20-year period was revealed in the 7 strains and in the ELD complex in general. A G-band idiogram of the normal mouse chromosomes was elaborated with numbering of regions and bands. By G-band analysis the following average composition of the ELD karyotypes was indicated: unchanged mouse chromosomes (30%,), marA, marB, minutes (11%), mar1—mar12, telocentric markers (10%), anonymous telocentric markers, often strongly rearranged (50%). Great differences were revealed among normal chromosomes and markers in their capacity to persist in the tumor stemline as structurally stable units. The presence of some unchanged chromosomes in most cells of all 7 strains indicated that they involved selective advantage. Evidently chromosomal evolution has operated by preferably maintaining certain elements unchanged and by submitting other elements to heavy rearrangement. The C-band analysis showed that, in addition to the centromeric C-heterochromatin present in all chromosomes, some had interstitial C-bands, as marA in all strains, mar5 and mar7 in specific single strains, and others in numbers varying among the strains.

Experimental elimination and recovery of double minute chromosomes in malignant cell populations.

The influence of the environment on the frequency of cells with double minute chromosomes (dms) was studied in the SEWA mouse ascites tumor. Under normal in vivo conditions, this tumor contained about 90%., cells with one or more dms. Explantation in vitro resulted in a decrease of the dms carrying cells, which after 100 days only amounted to 5% of the population. Following reimplantation in vivo. the original proportion of these cells was restored after an interval of 170 days. A mathematical model is presented describing these changes as selective phenomena. In the stock tumor almost all minute chromosomes were C-band negative. In the experimental populations. both in vivo and in vitro, another type of minute chromosomes appeared which were C-band positive. Also by other morphologic and functional criteria they were shown to be small chromosomes and not dms. In the present paper they have been referred to as microchromosomes. The fact that dms have been found only in malignant cells, and the correlation established in the present experiments between presence of dms and in vivo environment, led to speculations concerning their function. One possibility is that they involve amplification of genic material of significance to tumor development.

Have double minutes functioning centromeres?

Normally, double minute (DM) chromosomes are extremely difficult to observe at other mitotic stages than prophase and metaphase. In a certain clonal subline of the mouse ascites tumor SEWA, however, the DMs were unusually distinct and could be followed through all mitotic stages. Their behavior was extraordinary: During metaphase-anaphase they exhibited no response whatever towards the spindle forces, and it was concluded that the DMs were without functioning centromere. Instead, at metaphase they were attached to or enclosed by the nucleolar matter persisting around the chromosome ends, and at anaphase they were transported to the poles together with the nucleolar matter, sticking to the daughter chromosomes. The daughter halves of each DM were usually carried to the same pole. Thus most of the DMs, in spite of their lack of detectable centromeric activity, were included into the telophase nuclei. Some micronuclei were found, however, probably leading to the loss of a proportion of the DMs. The content of DMs in a certain tumor cell population therefore must depend on the balance between this loss and the gain due to a positive selection value of cells containing DMs.

A hexaploid Ehrlich-Lettré mouse ascites tumor line with low sensitivity to colchicine.

A hexaploid mouse ascites tumor, ELH, derivative of the hyperdiploid Ehrlich-Lettre tumor (ELD), has been studied from different view-points. It originated as an in vivo implant of the hyper-triploid in vitro strain ELDA, which in its turn developed from an in vivo strain of the original in vitro explant in 1964 of the ELD Lund strain. Chromosome number distribution and behavior of different marker chromosomes were followed during the prehistory of ELH and through serial in vivo and in vitro transplantation. Its C- and G-band patterns were analyzed in passages 264 in vivo and 170 in vitro, and its response to colchicine, in passages 105–115, in vivo. ELH was demonstrated to be about 10 times more resistant than ordinary hyperdiploid ELD strains both to the c-mitotic action and to the general toxicity of colchicine.

Large double minutes with ring-shape and rod-shape

High incidence of large double minutes (DM) was characteristic of one subline of the SEWA mouse ascites tumor. This subline, SEWA1R, was the one in which the new C-bandless chromosomes (CM) appeared. The incidence of large DM was associated with low incidence of DM per cell, low incidence of DM-carrying cells, and high incidence of CM-carrying cells. The large DM were either round or elongated (rod-shaped). Many of the former kind were suggestive of ring-shape and a few of the largest obviously constituted pairs of rings, free or interlocked. One configuration of 4 interlocked rings was evidently in its second mitotic generation; in the same cell another configuration of 2 pairs of interlocked rings had developed from 1 pair of free rings in the preceding generation. Possible relations between CM and large DM are considered.