ABSTRACT
roxburghii. Anticancer activity of MMR has been carried out on Ehrlich ascites carcinoma (EAC) cells with three different doses (20, 40 and 60 mg/kg/day) by observing different parameters such as tumor weight, survival time of EAC-bearing mice, growth inhibition of EAC cells, morphological changes and nuclear damage of EAC cells etc. whereas antioxidant activity was determined by measuring total antioxidant, DPPH free radical scavenging, ferrous reducing capacity assay. The extract showed highest anticancer activity at 60 mg/kg day¬-⻹(i.p.). It caused 81.4% (P<0.01) cells growth inhibition and reduced tumor burden significantly (78.5%; P<0.001) in comparison to control. It also increased life span of EAC-bearing mice significantly (73.5%; P<0.01). MMR treated EAC cells showed membrane blebbing, chromatin condensation, nuclear fragmentation (apoptotic feature) in Hoechst 33342 staining under fluorescence microscope. DNA fragmentation assay in agarose gel (1.5%) electrophoresis also rectified that it causes EAC cells death by apoptosis. MMR also exhibited moderate antioxidant properties in dose dependent manner. Thus, this plant can therefore be considering a resource for natural chemo-preventive drugs as well as a possible pharmaceutical supplement.
Subject(s)
Antineoplastic Agents/pharmacology , Antioxidants/pharmacology , Apoptosis/drug effects , Carcinoma, Ehrlich Tumor/drug therapy , Methanol/chemistry , Plant Extracts/pharmacology , Rubiaceae , Solvents/chemistry , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/isolation & purification , Antineoplastic Agents/toxicity , Antioxidants/chemistry , Antioxidants/isolation & purification , Antioxidants/toxicity , Biphenyl Compounds/chemistry , Carcinoma, Ehrlich Tumor/pathology , Cell Line, Tumor , DNA Fragmentation , Dose-Response Relationship, Drug , Lethal Dose 50 , Male , Mice , Phytotherapy , Picrates/chemistry , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Extracts/toxicity , Plant Leaves , Plants, Medicinal , Rubiaceae/chemistry , Time Factors , Tumor BurdenABSTRACT
Anticancer activities of p-menth-1-ene-4,7-diol (EC-1) isolated from Eucalyptus camaldulensis Dhnh. were studied on Ehrlich ascites carcinoma (EAC) cells by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. Anticancer activities also analyzed in EAC-bearing mice by assessment of cancer growth inhibition, changes in cancer volume, changes in life span, and hematological parameters. Apoptosis was analyzed by fluorescence microscope, DNA fragmentation assay, and flow cytometry. The expression of apoptosis-related genes, Bcl-2, Bcl-X, PARP-1, p53, and Bax, were analyzed using polymerase chain reaction (PCR). EC-1 significantly inhibited proliferation of EAC cells in vivo and restored the altered hematological parameters of EAC-bearing mice. Cytological observation by fluorescence microscope showed apoptosis of EAC cells upon treatment with EC-1. Also, DNA fragmentation assay revealed EAC cells' apoptosis following EC-1 treatment. Increased mRNA expressions of p53 and Bax genes and negative expressions of Bcl-2 and Bcl-X were observed in cells treated with EC-1. These findings confirmed the induction of apoptosis by EC-1. In addition, MTT assay showed dose-dependent anticancer activity of EC-1 against EAC cell. Cell cycle analysis revealed that EC-1 treatment caused suppression of EAC cells at S phase. To conclude, EC-1 is a novel anticancer compound and showed antiproliferative and apoptotic activities in cellular and mice models.
