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. 2007 Jun;170(6):1964–1974. doi: 10.2353/ajpath.2007.060935

Figure 4.

Figure 4

Effect of KGF on VEGF-A expression in pancreatic cancer cells. A: Time course of VEGF-A induction. Q-PCR analysis showed significant increases in VEGF-A mRNA levels at 3 and 6 hours after the addition of 10 ng/ml rhKGF to MIA PaCa-2 cells. Each experiment was performed twice, and gene expression measurements were performed in triplicate. Bars represent the mean ± SE (*P = 0.038, **P = 0.041). B: Effects of exogenous KGF on VEGF-A protein levels. After the addition of rhKGF (0 to 100 ng/ml) to MIA PaCa-2 cells, VEGF-A levels in the culture supernatant were measured by ELISA and were found to be increased significantly in a dose-dependent manner. Results shown represent the mean ± SE of two separate experiments, each conducted in triplicate (*P = 0.034, **P = 0.018). C and D: Effects of engineered expression of KGF on VEGF-A expression. PANC-1-KGF and PANC-1-Mock cells were incubated in serum-free medium for 72 hours. KGF and VEGF-A mRNA levels were then determined by Q-PCR analysis. KGF (*P < 0.0001; C) and VEGF-A (*P = 0.004; D) mRNA levels in PANC-1-KGF were significantly higher than in PANC-1-Mock cells. Each experiment was performed twice, and each measurement was performed in triplicate. Bars represent the mean ± SE. E and F: Effects of sh-KGFR on VEGF-A expression. Western blot analysis showed that sh-KGFR inhibited the expression of KGFR protein levels in MIA PaCa-2 cells compared with sh-control (*P = 0.0006; E). Cells were treated with rhKGF (10 ng/ml) after being transfected with sh-KGFR or sh-control. sh-KGFR transfection was associated with significantly reduced VEGF-A protein levels in the conditioned medium after the addition of rhKGF (10 ng/ml) compared with sh-control (*P = 0.0016; F). Each measurement was performed in triplicate. Bars represent the mean ± SE. G and H: Effects of sh-KGF on VEGF-A expression. sh-KGF and sh-control were transiently transfected into KLM-1 cells. Q-PCR revealed that sh-KGF significantly reduced KGF and VEGF-A mRNA levels in KLM-1 cells compared with sh-control (*P = 0.0002, **P = 0.0046; G). Furthermore, sh-KGF significantly reduced VEGF-A protein levels in conditioned medium compared with sh-control (*P = 0.0016; H). Each experiment was performed twice, and each measurement was performed in triplicate. Bars represent the mean ± SE.