Illumina HiSeq 2000 paired end sequencing; Differential gene expression for A2780 human ovarian cancer cells when exposed to a new osmium-based anticancer agent
Differential gene expression for A2780 human ovarian cancer cells when exposed to a new osmium-based anticancer agent
A2780 ovarian cancer cells were obtained from ECACC (European Collection of Animal Cell Culture) and grown in RPMI-1640 medium supplemented with 10% (v/v) fetal calf serum, 1% (v/v) 2mL L-glutamine and 1% (v/v) penicillin (10 k units/mL)/streptomycin (10 mg/mL), at 310 K in a humidified atmosphere containing 5% CO2. Maintenance passages were done at ca. 80% confluency. Cells were seeded at 3 × 10^6 in P100 petri dishes and left to incubate for 24 h at 310 K in a humidified atmosphere containing 5% CO2. Stock solutions of the negative control and compound 1 were prepared in 5% DMSO, 10% saline, and 85% RPMI-1640 medium. Cells were exposed to compound 1 at a final concentration of 0.15 μM. After 4, 12, 24 and 48 h exposure, medium was removed from triplicate compound-exposed and control-exposed dishes, and cells were washed twice with PBS and collected using trypsin.  Between 5 × 10^6 and 1 × 10^7 cells in each sample were taken forward for RNA extraction. Using QIAshredder spin columns (Qiagen) the cells were lysed, as per manufacturers instructions. DNA was eliminated from each sample using gDNA spin columns, and the RNA was purified using RNeasy spin columns and buffer solutions (RNeasy plus mini kit, QIagen) as per the manufacturers instructions. RNA was collected in 70 μL RNase-free water and stored at -70 C. Libraries were prepped using Truseq by the Oxford Genomics Centre, UK
Experimental Factor: 24
sampling time point
Experimental Factor: negative control
compound