Data are presented as mean ± SD. (Figure 2) Figure 2 VEGF and MMP-2 mRNA levels in SW1990 and Capan-2 cells were detected by real time PCR. The extracted total RNA was reverse-transcribed into single-stranded cDNA, and real-time PCR was performed. Interleukin-6 (IL-6) markedly increased MMP-2 and VEGF mRNA expression in Capan-2 cells(P = 0.000, P = 0.000). AG490 significantly decreased MMP-2 and VEGF mRNA expression in SW1990 cells(P = 0.008, P = 0.000). β-actin was used as an endogenous control.
* P < 0.01, versus Capan-2 cell group; #P < 0.01, versus SW1990 cell group. Effects of AG490 Selleckchem Adavosertib and IL-6 on INCB024360 concentration p-Stat3 protein expression in pancreatic cancer cells Immunocytochemical staining showed that p-Stat3 was mainly expressed in the nucleus and weakly expressed in the cytoplasm of SW1990
and Capan-2 cells. Treatment with 20 μM/L AG490 in SW1990 cells for 24 hours markedly decreased the intensity of p-Stat3 expression. Treatment with 100 ng/ml IL-6 in Capan-2 cells for 24 hours significantly increased the intensity of p-Stat3 expression. (Figure 3) Figure 3 p-Stat3 protein expression was detected by immunocytochemistry. Immunocytochemical staining showed that p-Stat3 was mainly expressed in the nucleus and weakly expressed in the cytoplasm of SW990 cells and Capan-2 cells. Expression of p-Stat3 protein in Capan-2 cells (A) and SW1990 cells (C). After treatment with interleukin-6 (IL-6) for 24 hours on IWR-1 in vivo Capan-2 cells (B), we observed that the intensity of p-Stat3 expression SPTLC1 increased(P = 0.012). After treatment with AG490 for 24 hours on SW1990 cells (D), we observed that the intensity of p-Stat3 expression decreased (P = 0.006) (original magnification, ×400). (E) Integrated optical density of every group. Bars indicate mean ± SD. * P < 0.01, versus Capan-2 cell group; # P < 0.01, versus SW1990 cell group. Effects of AG490 and IL-6 on p-Stat3, VEGF and MMP-2 protein levels in pancreatic cancer cells We used western blotting to examine the effects of AG490 and IL-6 on p-Stat3, VEGF, and MMP-2 protein levels of SW1990 and Capan-2
cells. AG490 did not affect total Stat3 protein levels in SW1990 cells after treatment with 20 μM/L AG490 for 24 hours but did suppress p-Stat3, VEGF, and MMP-2 protein levels. Treatment of Capan-2 cells with 100 ng/ml IL-6 for 24 hours increased p-Stat3, VEGF, and MMP-2 protein expression levels significantly. (Figure 4) Figure 4 Stat3, p-Stat3, MMP-2 and VEGF protein expression in SW1990 and Capan-2 cells were detected by Western blotting. Protein samples extracted from SW1990 and Capan-2 cells treated for 24 hours with AG490 and interleukin-6 (IL-6), respectively, were subjected to western blotting for Stat3, p-Stat3, MMP-2, VEGF and β-actin proteins. AG490 and IL-6 did not affect total Stat3 protein levels. AG490 decreased p-Stat3, MMP-2 and VEGF protein expression in SW1990 cells(P = 0.010, P = 0.000, P = 0.009).