FIG. 8.
COX2 and MMP9 gene expression. (A) HCT116 cell line. Cells were seeded (1,500,000 per well) in triplicate in 500 μL growing media and incubated for 24 h at 37°C in a humidified 5% CO2–95% air atmosphere. Cells were treated with 50 ng/mL TNFα overnight and then treated with C. sativa C2F or F7 added 5 h before RNA extraction. (B) Colon biopsies. C2F and F7 were added overnight to four ulcerative colitis patients and one Crohn's disease patient biopsies at concentrations of 0.2 and 0.07 mg/mL, respectively. RNA was extracted and reverse transcribed, and values of the steady-state level of gene transcripts were determined as the ratio between the target gene (COX2 or MMP9) and a reference gene (GAPDH), and that of treatment versus no treatment (NT) cells, using the 2ΔΔCt method. The experiment was performed in three biological replicates, with three technical repeats for each (n=3). SE was calculated for three biological replicates for each examined treatment. Different letters above bars indicate statistically significant differences between means by one-way analysis of variance with Tukey's HSD (p<0.01). COX2, cyclooxygenase-2; MMP9, metalloproteinase-9.