Article Figures & Data
Figures
- Figure 1.
Protein expression of Aurora-2 kinase in pancreatic cancer cell lines. Western blot analysis indicates an up-regulation of 3–4-fold of Aurora-2 in nine different pancreatic cancer cell lines (AsPC-1, BxPC-3, Capan-1, CFPAC-1, HPAF II, Mia PaCa-2, Mutj, Panc-1, and SU.86.86) in comparison with normal diploid foreskin fibroblast cells (Forf). β-actin served as an internal control.
- Figure 2.
Aurora-2 kinase expression in pancreatic patient tumors. Real-time PCR of pancreatic patient tumor samples indicates an up-regulation of Aurora-2 mRNA expression in samples 1, 4, 6, 8, and 9 taken directly from patients compared with normal pancreas tissue (N). Normalized ratio is obtained from pancreatic tumor tissue versus normal tissue.
- Figure 3.
Immunohistochemistry of pancreatic tissue array. A, an array of paraffin-embedded pancreatic tumor samples. B, a specific pancreatic tumor sample at higher magnification from the array indicates a high level of Aurora-2 protein expression.
- Figure 4.
Inhibition of Aurora-2 using antisense oligonucleotides. A, RT-PCR of a human pancreatic tumor cell line (Mia PaCa-2) treated with antisense Aurora-2 at 25, 50, 100, 200, and 400 nm concentrations. Antisense-treated cells indicate a significant inhibition of Aurora-2 expression at 200 nm concentration compared with the scrambled oligonucleotides. B, RT-PCR of time course study of Mia PaCa-2 cells treated with Aurora-2 antisense indicates inhibition of Aurora-2 expression as early as 6 h.
- Figure 5.
Inhibiting Aurora-2 arrests cell cycle. A, time-dependent study showing Western blot analysis of protein extracts from pancreatic cell line (Mia PaCa-2) treated with antisense oligonucleotides shows inhibition of Aurora-2 expression at 24 and 48 h. B, following inhibition of Aurora-2 expression using antisense oligonucleotides, the flow analysis indicates a fraction of cells in sub-G1 phase of the cell cycle in the antisense-treated cells compared with the untreated control, indication of cells undergoing apoptosis.
- Figure 6.
Inhibiting Aurora-2 expression induces caspase-3 activation. Mia PaCa-2 cells treated with Aurora-2 antisense show an induction of caspase-3 activity at 24 and 48 h. The untreated cells indicate a baseline of caspase-3 activity at 100% at 24 and 48 h. The antisense scrambled cells (AS Scramble) indicate an increase in caspase-3 activity at 24 h (9%) and 48 h (80%) compared with the untreated control cells. However, the antisense-treated (AS Treated) cells show a significant increase in apoptosis compared with the untreated and the antisense scrambled controls. The antisense-treated cells indicate a 20% increase in caspase-3 activity at 24 h and 90% increase at 48 h compared with the scrambled antisense controls.
Volume 3, Issue 4
