1. Pancreatic cancer cells selectively stimulate islet β cells to secrete amylin 
Xianzhong Ding, Peter R. Flatt, Johan Permert, Thomas E. Adrian 
Gastroenterology 
Volume 114, Issue 1, Pages (January 1998)
DOI: /S (98)
Copyright © 1998 American Gastroenterological Association Terms and Conditions

2. Fig. 1
Media glucose concentrations after BRIN-BD11 cells (2) or PANC-1 cells (♢) were cultured alone or cocultured (○) for 42 hours. Glucose concentrations were not significantly different between the three groups at 6, 12, 18, 24, or 36 hours. However, glucose concentration was significantly decreased in the coculture group at 42 hours compared with both control groups. Values are expressed as means ± SEM for 6 separate experiments. ★P < 0.05 vs. both control groups.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

3. Fig. 2
(A) Time course of amylin secretion from BRIN-BD11 cells cocultured with PANC-1 cells. Amylin secretion was significantly increased from BRIN-BD11 cells after 18 or 24 hours of coculture (PANC-1; ▩) compared with BRIN-BD11 cells cultured alone (control; 2). (B) In contrast, insulin secretion was not influenced by coculture with PANC-1 cells. Both amylin and insulin secretion increased with time in both coculture and control wells. Values are expressed as means ± SEM for 6 separate experiments. ★★P < 0.01 and ★★★P < vs. control group at same time point.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Effect of different numbers of PANC-1 cells on (A) amylin and (B) insulin secretion from BRIN-BD11 cells. After 24 hours of coculture, amylin secretion was greater with 25,000 or 50,000 PANC-1 cells compared with 10,000 cancer cells. Insulin secretion was not changed by the number of PANC-1 cells in the coculture system. Values are expressed as means ± SEM for 4 separate experiments. ★★P < 0.01 vs. group of 10,000 PANC-1 cells.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Effect of PANC-1 and Colo 320 cells on intracellular (A) amylin and (B) insulin content. Intracellular amylin content decreased in BRIN-BD11 cells cocultured with PANC-1 cells for 24 hours compared with control values (★★★P < 0.001). In contrast, PANC-1 cells had no effect on intracellular insulin content. Coculture with Colo 320 had no effect on amylin or insulin content. Values are expressed as means ± SEM for 6 separate experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

6. Fig. 5
(A) Amylin secretion from BRIN-BD11 cells cocultured with HPAF cells markedly increased compared with control values over 24 hours (★★★P < 0.001), and (B) the BRIN-BD11 cell content of amylin showed a small but significant decrease with coculture (★P < 0.05). Both (C) insulin secretion and (D) content were unaffected by coculture with HPAF cells. Values are expressed as means ± SEM for 4 separate experiments.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Effect of PANC-1 cell–conditioned medium on (A) amylin secretion and (B) content and (C) insulin secretion and (D) content. PANC-1 cell–conditioned medium was prepared as described in Materials and Methods. BRIN-BD11 cells were plated into 12-well plates at a density of 5 × 104/well. The cells were allowed to attach overnight and were washed with PBS, then cultured in PANC-1 cell–conditioned medium or control RPMI medium for 24 hours. Amylin and insulin were measured by radioimmunoassay. Values are expressed as means ± SEM for 6 separate experiments. ★P < 0.005, ★★P < 0.01 vs. control group.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

8. Fig. 7
Effect of HPAF-conditioned medium on (A) amylin secretion and (B) content and (C) insulin secretion and (D) content. HPAF cell–conditioned medium was prepared as described in Materials and Methods. BRIN-BD11 cells were plated into 12-well plates at a density of 5 × 104/well. The cells were allowed to attach overnight, washed with PBS, and cultured in HPAF-conditioned medium or control RPMI medium for 24 hours. Amylin and insulin were measured by radioimmunoassay. Values are expressed as means ± SEM for 6 separate experiments. ★P < 0.05, ★★P < 0.01 vs. control group.
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions

9. Fig. 8
Effect of different fractions of PANC-1 cell–conditioned medium, separated by gel permeation chromatography on amylin and insulin secretion. Six crude fractions were eluted from the Superdex peptide HR10/30 column. BRIN-BD11 cells were cultured with each fraction for 24 hours, and amylin and insulin in the culture medium were measured by radioimmunoassay. The major amylin-releasing activity was eluted in the third fraction of PANC-1 cell–conditioned medium, which corresponds to the molecular weight of  ̃1500 (P < 0.01 vs. control).
Gastroenterology  , DOI: ( /S (98) )
Copyright © 1998 American Gastroenterological Association Terms and Conditions