Article Figures & Data
Figures
- Figure 1.
RON tyrosine kinase receptor is expressed in human and murine pancreatic cancer. A, immunoblot analysis of pancreatic cancer cell line lysate shows RON expression in five of seven human pancreatic cancer cell lines (ASPC-1, BxPC-3, Capan-1, HPAF-II, Hs766T) and five of five cell lines derived from PdxCre/LSL-KRASG12D mice, including primary pancreatic carcinoma (4964PDA, 5143PDA), liver metastatic lesions (4964LM, 5143LM), and PanIN lesions. Below the actin control band corresponding to the human cell lines, a longer exposure of the human cell lines immunoblot is seen to show the presence of the 150-kDa β-chain RON in Hs766T. B, the overall number of tissue samples expressing RON increased with severity of disease. The numbers above each bar graph represent the total number of evaluated samples possesing the indicated lesions on the X-axis. C, human pancreatic cancers were immunostained with a RON-specific antibody and the intense RON expression correlates with pancreatic cancer progression. Minimal expression is seen in normal pancreatic ductal epithelium with obvious expression present in PanIN 1B lesions that persists in later stage PanINs, invasive carcinoma, and in both lymph node (LN) and liver metastases. A series of RON-positive invasive carcinomas (n = 8) were also immunostained for the phosphorylated, active form of the RON receptor and showed that 100% of samples stained positive, with a single representative picture shown. This shows that total RON expression in pancreatic cancer specimens correlates with the presence of active, phosphorylated RON.
- Figure 2.
RON activation has no effect on proliferation in murine and human pancreatic cancer cell lines. Alamar Blue proliferation assays were done in 5143PDA (A), 5143LM (B), PanIN (C), and BxPC-3 cells (D). Baseline fluorescent measurements corresponding to cell proliferation were acquired at 0 h, followed by 12, 24, and 48 h post-HGFL administration. No statistical difference in proliferation was seen in any cell line from 0 to 48 h post-HGFL. P > 0.05 for all time points compared with untreated control.
- Figure 3.
RON activation increases migration, invasiveness, and apoptotic resistance in pancreatic cancer cells. A, pancreatic cancer cells showed a dose-dependent increase in migration. B, the ability to invade a Matrigel basement membrane in response to the RON ligand, HGFL. This ability to migrate in response to 100 ng/mL of HGFL was abrogated by preincubation with inhibitory RON antibody (C). D, incubating BxPC-3 cells with inhibitory RON antibody before treatment with 0.1 μmol/L gemcitabine resulted in a 32% increase in apoptosis. *, P < 0.05.
- Figure 4.
RON activation in pancreatic cancer cells results in the up-regulation of known oncogenic signaling pathways. Immunoblot analysis for total and phosph-Akt, phospho-MAPK, and IκB-α and IκB-β following exposure to HGFL in the murine PanIN cell line and in the human pancreatic cancer cell lines ASPC-1 and BxPC-3. A clear up-regulation of the active, phosphorylated forms of Akt and ERK are shown in all cell lines after 1 h of RON activation. However, no differences in IκB levels were detected post-HGFL compared with control.
Volume 67, Issue 13
