Different theories of carcinogenesis.

<p>The SMT postulates that mutations in oncogenes lead to cell-autonomous growth. Additional mutations are necessary for invasive growth (<b>A</b>). The tissue-based models assume that reciprocal communication between tumor and stromal cells can influence the phenotype of the tumor cells. The tissue-based models question the autonomy of the mutations in epithelial cells and the sovereignty of the tumor cells in determining the cancer phenotype (<b>B</b>). In the FBM, mutations in genes could show a normal phenotype under physiological conditions but can be activated by inflammation. Under inflammatory conditions, tumor and stromal cells interact to form a positive feedback loop. Additional stromal and inflammatory cells become attracted to the inflammatory micromilieu (<b>C</b>).</p

Normal activation of NF-κB and KRAS in an inflammatory environment.

<p>Negative feedback loop within the NF-κB pathway by SOCS and A20 counteracts the activated state of the epithelial cell (<b>A</b>). The non-physiological activation of KRAS impairs the negative feedback loop within the NF-κB pathway and leads to a perpetual inflammatory microenvironment (<b>B</b>).</p

Effect of treatment with GPRC5a-siRNA over 72 hours.

<p>A: Number of living cells in negative controls (NC; 1: Medium, 2: non-sense-siRNA), positive control (PC; eg5-siRNA) and knock-down with two different GPRC5a-siRNA (RAI3_A and RAI3_B). Statistically significant differences in t-test are marked with * for P-Value < 0.001 and ** for P-Value < 0.05; B: Western blot and quantitative real time PCR with RAI3 antibody confirms the successful knock-down using siRNA (for functional analysis the GPRC5A.1 siRNA was chosen) (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0170390#pone.0170390.s001" target="_blank">S1 Fig</a>). (n = 5).</p

Multivariate Cox proportional hazard model for different variables.

<p>Multivariate Cox proportional hazard model for different variables.</p

Univariate analysis of clinico pathological variables of the 376 patients with pancreatic cancer analyzed on the TMA.

<p>Univariate analysis of clinico pathological variables of the 376 patients with pancreatic cancer analyzed on the TMA.</p

Effect of siRNA based knock-down on the colony formation and migration of pancreatic cancer cell lines.

<p>A: Examples of the results of typical colony forming assays over 7 days after siRNA-treatment; B: Growth area compared to NC1 (statistically significant difference between RAI3 and NC2 is demonstrated by * for P<0.01 and ** for P<0.001 performed by t-test); C: Typical images of migrated cells in negative control (NC) and knock-down with GPRC5a siRNA (RAI3) in different cell lines; D: Number of migrated cells after 48 hour siRNA-treatment and subsequently 16 hours migration assay (* indicates statistically significant differences (P<0.05) in t-test between the negative control with non-sense-siRNA (NC) and the knock-down with GPRC5a-siRNA (RAI3)). (n = 3).</p

Bar graphs of the pre- and postoperative values for (A) HbA1c, (B) BMI, (C) Ca19-9 and (D) CEA with respect to deteriorated, stable and improved blood glucose homeostasis.

<p>Points: means; error bars: 95% CI; black error bars: preoperative values; broken error bars: postoperative values for each group; reference lines: cut-off/normal values; F- and p-values beneath each diagram: one-sided ANOVA for comparison between the three groups at each time point; and p-values within the diagram: two-sided T-test for paired samples for pre- and post-operative comparisons within the designated group.</p

Distribution of the categorical variables based on the dynamics of blood glucose homeostasis after partial pancreatectomy.

<p>Distribution of the categorical variables based on the dynamics of blood glucose homeostasis after partial pancreatectomy.</p

RAI3 immunohistochemistry as diagnostic and prognostic instrument.

<p>A: Definition and calculation of a diagnostic score for the presence of a pancreatic ductal adenocarcinoma (PDAC); B: Receiver Operating Characteristic (ROC) curve for the diagnostic score using for distinction of PDAC from Normal or Chronic Pancreatitis (CP) tissue (Sensitivity and specificity were calculated for two exemplary cut-off-points); C: Univariate Kaplan-Meier analysis (statistically significant differences between apical expression over 30% and less (p = 0.017) according to Log-Rank-test).</p

Expression of RAI3 in human tissue and cell lines.

<p>TMA—Immunochemistry with RAI3-Antibody in different types of pancreatic tissue. Presentation in 20-fold magnification of A-C: Normal; D: Chronic pancreatits (CP), E: Intraductal Papillary Mucinous Neoplasm (IPMN); F—H: Pancreatic Ductal Adenocarcinoma (PDAC). Three parameters were considered: RAI3-Intensity (I), RAI3-Expression (Ex) and Apical RAI3-Expression (aEx); I,J: Differential expression of RAI3 mRNA respective protein in common pancreatic cell lines and primary pancreatic cell lines measured by qRT-PCR (I) and Western Blot (J) with a conspicuously low expression in BxPc3 and PaCaDD165 in comparison to the other cell lines in the group. (n = 3).</p