Early prediction of persistent organ failure in patients with acute pancreatitis

s / Pancreatology 16 (2016) S1eS63 S35 Introduction: Pancreatic ductal adenocarcinoma (PDAC) is still one of the poorest prognoses cancers, lacking understanding. In PDAC, abnormal activity of Hedgehog (HH) pathway, inflammation(Nuclear factor-kappa B, NF-kB) and K-ras mutation were all involved in. Exploring the interaction of the three elements of PDAC is quite critical to uncover the mechanism of PDAC. Methods: K-ras gene-type, protein expression of Shh and nuclear protein expression of Gli1 and NF-kB of 32 PDAC tissues were detected, the correlation of the three factors were also analyzed. SW1990 were transfected with Gli1 cDNA;SW1990, Panc-1and BxPC-3 were treated with Shh, using untreated group as control, then we examined expression of Shh, Gli1 and NF-kB, cell proliferation and apoptosis changes; K-ras downstream p-/t-ERK1/2 and p-/t-AKT1 expression, as well as Ras activity were tested to estimate K-ras activation. And, the correlation between HH pathway and NF-kB in different K-ras gene-type cells were analyzed separately. Then K-ras expression was knocked-down by si-Kras, experiments above were done again. At last, mutant K-ras cDNA was transfected into K-ras wild-type cells, Shh, Gli1 and NF-kB mRNA expression changes were detected after Shh stimulation. Results: Positive correlation of HH pathway and NF-kB was significant in K-ras mutant PDAC tissues. In K-ras mutant PDAC cells, HH activation could promote NF-kB activation, accelerate cell proliferation, and inhibit apoptosis, up-regulate p-ERK1/2 and p-AKT1, and increase active-Ras expression. When K-ras expression was down-regulated by si-Kras, all the effects above were all weakened. But in wild-type K-ras cells, little significant change happened under such circumstances. But when they transfectedwithmutant K-ras cDNA, themRNA expression of Shh, Gli1 and NF-kB were all increased significantly by Shh stimulation. Conclusions: HH pathway induced NF-kB activation was based on mutant K-ras gene-type in PDAC, and K-ras activity change is the critical molecular mechanism behind them.