Acanthus ilicifolius plant extract prevents DNA alterations in a transplantable Ehrlich ascites carcinoma-bearing murine model

AIM： To investigate the chemopreventive efficacy of the Indian medicinal plant Acanthus ilicifolius L Acanthaceae in a transplantable Ehrlich ascites carcinoma （EAC）- bearing murine model.METHODS： Male Swiss albino mice were divided into four groups： Group A was the untreated normal control; Group B was the EAC control mice group that received serial, intraperitoneal （ip） inoculations of rapidly proliferating 2 × 10^5 viable EAC cells in 0.2 mL of sterile phosphate buffered saline; Group C was the plant extract-treated group that received the aqueous leaf extract （ALE） of the plant at a dose of 2.5 mg/kg body weight by single ip injections, once daily for 10, 20 and 30 consecutive days following tumour inoculation （ALE control）; and Group D was the EAC ＋ ALE- treatment group. The chemopreventive potential of the ALE was evaluated in a murine model by studying various biological parameters and genotoxic markers, such as tumour cell count, mean survival of the animals, haematological indices, hepatocellular histology, immunohistochemical expression of liver metallothionein （MT） protein, sister-chromatid exchanges （SCEs）, and DNA alterations.RESULTS： Treatment of the EAC-bearing mice with the ALE significantly （P 〈 0.001） reduced viable tumour cell count by 68.34% （228.7 × 10^6 ± 0.53） when compared to EAC control mice （72.4 × 10^6 ± 0.49）, and restored body and organ weights almost to the normal values. ALE administration also increased （P 〈 0.001） mean survival of the hosts from 35 ± 3.46 d in EAC control mice to 83 ± 2.69 d in EAC ＋ ALE-treated mice. Haematological indices also showed marked improvement with administration of ALE in EAC-bearing animals. There was a significant increase in RBC count （P 〈 0.001）, hemoglobin percent （P 〈 0.001）, and haematocrit value （P 〈 0.001） from 4.3 ± 0.12, 6.4 ± 0.93, and 17.63 ± 0.72 respectively in EAC control mice to 7.1 ± 0.13, 12.1 ± 0.77, and 30.23 ± 0.57 respectively in EAC ＋ ALE-treated group, along with concurrent decrement （P 〈 0.001） in WBC count from 18.8 ± 0.54 in EAC control to 8.4 ± 0.71 in EAC ＋ ALE. Furthermore, treatment with ALE substantially improved hepatocellular architecture and no noticeable neoplastic lesions or foci of cellular alteration were observed. Daily administration of the ALE was found to limit liver MT expression, an important marker of cell proliferation with concomitant reduction in MT immunoreactivity （62.25 ± 2.58 vs 86.24 ± 5.69, P 〈 0.01）. ALE was also potentially effective in reducing （P 〈 0.001） the frequency of SCEs from 14.94 ± 2.14 in EAC control to 5.12 ± 1.16 in EAC ＋ ALE-treated group. Finally, in comparison to the EAC control, ALE was able to suppress in vivo DNA damage by abating the generations of＇tailed＇ DNA by 53.59% （98.65 ± 2.31 vs 45.06 ± 1.14, P 〈 0.001）, and DNA single-strand breaks （SSBs） by 38.53% （3.14 ± 0.31 vs 1.93 ± 0.23, P 〈 0.01） in EAC-bearing murine liver.CONCLUSION： Our data indicate that, ALE is beneficial in restoring haematological and hepatic histological profiles and in lengthening the survival of the animals against the proliferation of ascites tumour in vivo. Finally, the chemopreventive efficacy of the ALE is manifested in limiting MT expression and in preventing DNA alterations in murine liver. The promising results of this study suggest further investigation into the chemopreventive mechanisms of the medicinal plant A. ilicifolius in vivo and in vitro.

Antitumorigenic Potential of Diallyl Sulfide in Ehrlich Ascites Tumor Bearing Mice

Objective To study the effects of diallyl sulfide (DAS), an organosulfur compoundpresent in garlic (Allium sativum), on the life span of ehrlich ascites (EA) tumor bearingSwiss albino mice, cytotoxicity and angiogenesis. Methods EA tumor cells weremaintained by serial transplantation in peritoneal cavity of male Swiss albino mice. EAtumor cells were inoculated at concentrations of 1× 106 EA cells, 2.5× 106 EA cells and5× 106 EA cells. DAS was given in 0.2 ml normal saline i. p., daily for seven days followedone hour later by inoculation with EA cells in respective groups. Results The resultsrevealed that administration of DAS increased the life span of EA tumor bearing animals bymore than 25 percent. A significant dose dependant cytotoxic response of DAS was alsoobserved on EA tumor cells. DAS was also found to inhibit the angiogenesis in EA tumorbearing mice in a dose dependent manner. Conclusion It is suggested that DAS may exertits anticarcinogenic effects by more than one mechanism and is a useful chemopreventiveand chemotherapeutic agent.

Caffeic acid and protocatechuic acid modulate Nrf2 and inhibit Ehrlich ascites carcinomas in mice

Objective:To assess the nuclear factor-erythroid 2-related factor-2(Nrf2)modulatory effect of caffeic acid and protocatechuic acid and determine the anti-tumor activity of these phenolic compounds against Ehrlich ascites carcinoma growth in mice.Methods:Antioxidant activity of protocatechuic acid and caffeic acid was assessed using ferric reducing antioxidant power(FRAP)and 2,2-diphenyl-1-picrylhydrazyl(DPPH).Nrf2 activation potential of phenolic compounds was tested by quantitative realtime polymerase chain reaction,and luciferase complementation reporter assays.In vivo efficacy was tested using the Ehrlich ascites carcinoma model.Results:FRAP and DPPH radical scavenging assays showed that caffeic acid and protocatechuic acid were more potent compared with cinnamic acid and benzoic acid.Luciferase complementation reporter assays identified caffeic acid and protocatechuic acid as the activators of Nrf2.Both caffeic acid and protocatechuic acid upregulated the expression of Nrf2 target genes heme oxygenase-1(HO-1),glutamate-cysteine ligase catalytic subunit(GCLC),and glutamate-cysteine ligase modifier subunit(GCLM)and the activity of NAD(P)H:quinone oxidoreductase 1(NQO1)when tested on HCT-116 cells using a cell-based assay system at 9 h.In addition,intraperitoneal administration of caffeic acid and protocatechuic acid to Ehrlich ascites carcinoma bearing mice suppressed tumor growth and angiogenesis.Conclusions:Caffeic acid and protocatechuic acid can modulate Nrf2 and inhibit Ehrlich ascites carcinoma cells.

THE INHIBITORY EFFECT OF EXTRACT OF CAMELLIA SINENSIS AND EXTRACT OF CAMELLIA PTILOPHYLLA CHANG ON DNA POLYMERASE OF EHRLICH ASCITES CARCINOMA CELLS

Objective: To detect the effect of extract of Camellia Sinensis (ECS) and extract of Camellia Ptilophylla Chang (ECPC) on DNA polymerase (Pol) of Ehrlich ascites tumor cells Methods: Referring to the method of K Ono, Pol was extracted from Ehrlich ascites tumor cells in mice Pol α, β, and γ were separated by phosphocellulose column chromatography and were identified The effect of ECPC and ECS on Pol was studied Results: ECPC and ECS were shown to inhibit the activity of Pol α, β, and γ IC 50 values of ECS on Pol α , β, and γ were 10 2μg/ml, 9 9μg/ml and 28 9μg/ml respectively IC 50 values of ECPC on Pol α, Pol β and Pol γ were 5 6μg/ml, 15μg/ml and 14 7μg/ml respectively The modes of inhibition of ECPC on Pol α, Pol β and Pol γ were noncompetitive with respect to template DNA The Ki values of ECPC on Pol α , β, and γ were 2 68±0 12μg/ml, 2 24±0 12μg/ml , 2 56±0 18μg/ml Conclusion: ECPC and ECS were shown to have inhibitory effect on DNA polymerase of tumor cells The mode of inhibition of ECPC on Pol α, Pol β and Pol γ were noncompetitive with respect to template DNA

Anti-proliferative and apoptotic efficacy of diallyl disulfide onEhrlich ascites carcinoma

Aim:This study was conducted to assess the in vivo and in vitro anti-tumor effects of diallyl disulfi de(DADS)against Ehrlich ascites carcinoma(EAC)and to suggest its probable mechanism of action.Methods:EAC was induced in female mice by intraperitoneal injection of EAC-cells from stock mice.EAC-bearing mice were orally treated with 100 mg/kg body weight for 2 weeks beginning from the 1st day of EAC intraperitoneal transplantation.Cytotoxicity effects of DADS against EAC-cells in vitro were investigated at different concentrations(0,6.25,12.5,25,50,and 100μg/mL)of DADS using trypan blue exclusion assay.Results:Data from this study exhibited a signifi cant decrease in EAC-aliquot volume as well as total and alive EAC-cell number and a marked increase in dead EAC-cell number and percent in EAC-bearing mice treated with DADS as compared with EAC-bearing control.These changes were consistent with increased number of cells which exhibited phenotypic apoptotic signs marked by a decrease in the expression of anti-apoptotic protein Bcl-2,an increase of pro-apoptotic and cell cycle arrest mediator p53 and an elevation of DNA fragmenting indicator terminal deoxynucleotidyl transferase in EAC-bearing mice treated with DADS.In addition,the tumor marker sialic acid level was markedly decreased in plasma and Ehrlich ascites in EAC-bearing mice treated with DADS.In vitro,DADS also produced anti-proliferative and anti-tumor cytotoxic potentials against EAC.Conclusion:DADS may have anti-cancer effects which may be mediated via modulation of apoptosis and cell cycle arrest.

A Study in vitro on the Synergetic Cytocidal Effect ofAscorbic Acid and Mitomycin C against the Ehrlich As-cites Tumor Cells in Culture

The synergetic cytocidal effect of ascorbic acid and mitomycin C on Ehrlich as-cites tumor cells in culture was studied. The chain scission of DNA by Cu(Ⅱ)mitomycin C/ascorbic acid system,and the chemical kinetics of the Cu(Ⅱ) catalyzed aerobic oxidation of ascorbic acid in the presence of mitomycin C were also discussed. Experimental results showed that ascorbic acid and mitomycin C were synergetic in destroying Ehrlich ascites tumor cells.The effect is related to the breaking action of the Cu(Ⅱ)mitomycin C/ascorbic acid system on the DNA chain, and to the quantity of the system produced ·OH, and the speed of production which are both directly proportional to the concentration of ascorbic aci.These results imply that the ·OH produced in the aerobic oxidation of ascorbic acid,and·OH induced scission of DNA chain, are important factors to he synergetic cytocidal effect of ascorbic acid and mitomycin C on Ehrlich ascites tumor cells.

THE SENSITIVITY OF EHRLICH ASCITIC CARCINOMA CELLS TO THE EMPERIPOLESIS OF THYMUS LYMPHOCYTES

In mixed lymphocyte-tumor all culture in vitro, the ability of thymus lymphocytes adhering to tumor cells was found to be essentially consistent with its emperipolesis index, though not synchronous. Tumor cells at different progressive stages and in the mixed cultures with or without Con A stimulation also varied in the sensitivity to lymphocyte adhesion and emperipolesis. Tumor adhesiveness anl emperipolesis of theymus lymphocytes and their PNA-, PNA+ subgroups were shown to be different significantly from splenic lymphocytes. Thymosin exhibited certain promotive effect on lymphocyte-tumor adhesion and emperipolesis.

Antitumor Actinity of Puqietinone,a Novel Alkaloid fromthe Bulbs of Fritillaria puqiensis

从蒲圻贝母中分得的一种新生物碱蒲贝酮碱，按一定剂量灌胃给药，显示了强的抗小鼠艾氏腹水癌（ＥＡＳ，实体型），宫颈癌（Ｕ＿（１４），实体型）及肝癌（ＨｅＰＡ，实体型）的活性。

Production,characterization and biological activities of acidic exopolysaccharide from marine Bacillus amyloliquefaciens 3MS 2017

Objective: To evaluate in-vitro antioxidant, anti-inflammatory and antitumor abilities against human breast adenocarcinoma(MCF7) and human prostate cancer(PC3) as well as the suppressor effect of bacterial exopolysaccharide(BAEPS) on Ehrlich ascites carcinoma(EAC). Methods: In-vitro antioxidants characters of BAEPS were determined using various methods, while anti-inflammatory activity was estimated against cyclooxygenase(COX-1 and COX-2). In-vitro study, anticancer against MCF7 and PC3 were assessed by the mitochondrial dependent reduction of yellow MTT. In in-vivo study against EAC progression, mice were inoculated with EAC cells and then were orally administered BAEPS at 200 mg/kg after 24 h(equals to 0.10 of determined LD50)/10 d. Results: BAEPS was acidic exopolysaccharide contained uronic acid(12.3%) and sulfate(22.8%) with constitution of glucose, galactose and glucuronic acid in a molar ratio1.6: 1.0: 0.9, respectively, with a molecular mass of 3.76伊104 g/mol. BAEPS appeared potent antioxidant characters as free radical scavenging, oxygen reactive species scavenging and metal chelation, while its reducing power was low. BAEPS showed selective anti-inflammatory activity against COX-2 than COX-1, COX-2 selective. BAEPS exhibited potent and selective effect to breast cell cancer MCF7, the death percentage was 65.20% with IC_(50)=70 μg/m L and IC_(90)=127.40 μg/m L. BAEPS decreased counted viable EAC cells and induced non-viable cells. BAEPS improved all assessed hematological parameters. These improvements were reflected in the increasing median survival time and significant increment(P<0.05) in life span. Conclusions: BAEPS has anti-tumor activity with a good margin of safety. The anti-tumor activity of BAEPS may be due to its content from sulfated groups and uronic acids and they have antioxidant and anti-inflammatory properties.

Anticancer activity of Gloriosa superba Linn in EAC tumor-bearing Balb/c mice

Cancer is a pathogenic condition due to abnormal growth of cells,which is one of the leading reasons for the death of majority of the population throughout the world.Pharmacotherapy associated with the majority of the adverse effects such as cardiotoxicity,hepatotoxicity,nephrotoxicity that suggests identifying new drug which would be safer than well-established chemotherapy.There is a need to get a promising line for research over various diseases including cancer.Gloriosa superba Linn has a long history of use in folk medicine including wounds,hemorrhoids,kidney problems,and tumors.The present study explores the ethanolic extract of Gloriosa superba Linn as the anti-cancer agent using in vitro MTT assay and in vivo Ehrlich’s ascites carcinoma(EAC)bearing Balb/c mice models.Further,the study reports the IC50 value of the DPPH was 1.21μg/mL,and the IC50 of MTT assay against Hela and MCF7 was found to be 148.50μg/mL and 147.23μg/mL respectively.For,in vivo study,animals were divided into five groups(n=6)experimented for 14 days.The tumor was induced by intraperitoneal transplantation of Ehrlich’s ascites carcinoma cells in Balb/c mice.The normal and control group received normal saline.After 24 hours of Ehrlich’s Ascites Carcinoma transplantation,treatment was started by injecting Cyclophosphamide(50 mg/kg)intraperitoneal whereas Gloriosa superba Linn extract(5 mg/kg&10 mg/kg)was administrated orally.On the 15th day,mice were sacrificed to estimate hematology,biochemical estimation,and histopathology.The extract showed a significant decrease(P<0.001)in body weight in the treatment group compare to the control group further there was amelioration in hematology,biochemical estimation,and histopathology.From the result,it was concluded that the extract has a potent antitumor activity.

In vitro cytotoxic studies of red algae Portieria hornemannii and Spyridia fusiformis against Dalton’s lymphoma ascite and Ehrlich ascite carcinoma cell lines

Objective:To study the in vitro cytotoxic activities of methanol extract of Portieria hornemannii(P.hornemannii)and Spyridia fusiformis(S.fusiformis)using Dalton’s lymphoma ascite and Ehrlich ascite carcinoma cell lines.Methods:The effect of cytotoxicity of P.hornemannii and S.fusiformis was evaluated with the concentrations(100 to 200μg/mL)and assessed for the antitumour activity vs.the selected cell lines using Trypan blue assay.Results:The methanol extracts of P.hornemannii and S.fusiformis showed potent cytotoxic activity with IC_(50)values of(209.00±0.05)μg/mL and(190.00±0.05)μg/mL against the Dalton’s lymphoma ascite cell line and IC_(50)values of(190.00±0.05)μg/mL and(182.00±0.05)μg/mL against the Ehrlich ascite carcinoma cell line respectively.In vitro cytotoxicity against the tested cancer cell lines showed strong activity by the abnormal activities of algal residue in the normal cells.Conclusions:The methanol solvent residue of red algae(P.hornemannii and S.fusiformis)could be a good candidate.It would be a novel marine resource as a antitumor medicine demonstrated by cytotoxic studies that the above marine algae can be a potential candidate sources as antitumor drugs.

Phytochemical Profiling and Bioactivity of A Mangrove Plant,Sonneratia apetala,from Odisha Coast of India

Objective： To test the antioxidant, antidiabetic, anticancer and antibacterial activities along with phytochemicals of Sonneratia apetala Buch.-Ham. Methods： The antibacterial activity was determined by agar well diffusion method. The antioxidant activity was determined by standard assay. The antidiabetic activity was evaluated by α-glucosidase inhibition assay and in vivo anticancer property was determined against Ehrlich ascites carcinoma （EAC） cells in Swiss Albino mice. Further partial characterization of the methanol extracts was carried out by thin layer chromatography, high performance liquid chromatography, 1H nuclear magnetic resonance spectroscopy and Fourier transform-infra red spectrum spectral analysis. Results： Four solvent extracts （acetone, ethanol, methanol and aqueous） of leaf and bark possess strong antioxidant properties. In vivo anticancer activity of methanol extract leaf indicated positive activity showing 34% inhibition against EAC cells in Swiss Albino mice. All extracts exhibited a-glucosidase inhibitory activity in a dose-dependent manner indicating presence of promising antidiabetic properties. The extracts possess strong antibacterial activity against the selected pathogenic bacteria （minimal inhibitory concentration ranging from 1.25-5.00 mg/mL）. The partial characterization of the methanol extracts of leaf and bark revealed the presence of phenolics as the lead compound responsible for studied bioactivities of the plant extracts. Conclusion： Sonneratia apetala extracts have potent antibacterial, antioxidant, antidiabetic and anticancer properties which can be further exploited for its pharmaceutical applications.

In vivo anticancer activity of vanillin,benzophenone and acetophenone thiosemicarbazones on Swiss albino mice

Objective:To study the anticancer activities of three schiff bases viz.vanillin thiosemicarbazone,benzophenone thiosemicarbazone and acetophenone thiosemicarbazone against Ehrlich ascites carcinoma(EAC)cells in Swiss albino mice.Methods:Synthesized compounds have administrated into the intraperitoneal cavity of the EAC inoculated mice at two doses.The anticancer activities have studied by monitoring the parameters such as cell growth inhibition,tumor weight measurement,survival time of EAC bearing mice as well as the changes in depleted hematological parameters due to tumorgenesis.All such data have been compared with those of a known standard drug bleomycin at the dose of 0.3 mg/kg(i.p.).Results:It has been found that these bases enhanced life span,reduced average tumor weight and inhibited tumor cell growth of EAC cell bearing mice remarkably.The results were similar in potency to those obtained with bleomycin.It was also found that the depleted hematological parameters(red blood count,white blood count and haemoglobin content)were found to be restored gradually towards normal within few weeks after ceasing the treatment.Conclusions:The compounds can be primarily considered more or less as potent anticancer agents.