The present study was designed to evaluate the cytotoxic effect of
The present study was designed to evaluate the cytotoxic effect of methanol extract of aerial parts including stems, leaves and twigs of and purified continentalic acid isolated from this extract against a panel of human cancer cell lines of varied tissues. hypoglycemic activity of the roots of and evaluation of its immunomodulatory activity has already been reported by our research group. Continentalic acid has been reported for its analgesic activity, growth inhibition and apoptosis induction, antibacterial activity and antiinflammatory activity[24,25,26]. Earlier, continentalic acid has been reported to show moderate cytotoxicity against L1210, K562 and LLC tumor cell lines using MTT assay. So far, the cytotoxicity of has notbeen reported. Herein, we statement the evaluation of cytotoxic effect of crude methanol extract from aerial parts of and purified continentalic acid (CA, fig. 1) isolated from this extract against a panel of five human malignancy cell lines by sulphorhodamine B assay to explore their possible role in malignancy therapy. Open in a separate windows Fig. 1 Structure of continentalic acid. TLC was performed on 0.25 mm silica gel 60 F254 plates. Silica gel 60-120 mesh was utilized for column chromatography. HPLC analysis was performed using Agilent 1100 series. LC conditions employed were as follows: C8 column reversed-phase (E-Merck, 2504.0 mm, 5 m particle size) maintained at 30o, quaternary pump, photodiode array detector, water-acetonitrile (1:4, v/v) isocratic mobile phase at a circulation rate of 0.5 ml/min using automatic sample injection module. The aerial parts of herb were collected from khillanmarg area of Kashmir (Jammu and Kashmir), India after proper identification and authentication of herb by department of Bioscience and Biotechnology, Banasthali University or college, Rajasthan, India. (Sample no. BV 168). Reference specimen has been preserved in our laboratory. These herb parts were dried in shade and pulverized separately in a mechanical grinder, exceeded through a 40 mesh sieve, and stored in a closed vessel. This crushed material (680 gms) was extracted with real methanol through percolator for 72 h. The suspension was filtered and the filtrate was concentrated under reduced pressure to yield 87.95 gm of methanol free semisolid mass. Column chromatography of this extract using n-hexane:ethyl Rivaroxaban cost acetate gradient as eluting answer yielded real continentalic acid Pten as reported earlier. The human malignancy cell lines used in present study were procured from National Malignancy Institute, Frederick, U.S.A. Cells were grown in tissue culture flasks in total growth medium (RPMI-1640 Rivaroxaban cost medium with 2 mM glutamine, 100 g/ml streptomycin, pH 7.2, sterilized by filtration and supplemented Rivaroxaban cost with 10% fetal calf serum and 100 models/ml penicillin before use) at 37 in an atmosphere of 5% CO2 and 90% relative humidity in a carbon dioxide incubator. These cells at sub confluent stage, when the cells were 60-70% confluent, were harvested from your flask by treatment with trypsin (0.5% in PBS containing 0.02% EDTA). Cells with viability of more than 98%, as determined by trypan blue exclusion, were utilized for the present study. The cell suspension of the required cell density (1105 cells/ml) was prepared in complete growth medium made up of gentamicin (50 g/ml) for determination of cytotoxicity. Stock solutions of 410-2 M of continentalic acid and 1000 g/ml of extract were prepared in DMSO. DMSO was used in such a way that final concentration in the dilution was less than 1%. However the control cells, which were not given any treatment, were having equivalent amounts of DMSO, to nullify the effect. The stock solutions were serially diluted with total growth medium made up of 50 g/ml of gentamicin to obtain working test solutions of required concentrations and were stored at -20 until use. cytotoxicity of test materials was evaluated.