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2001 Abstract: 1793 A 2026 D Peptide as the Putative Pancreatic Cancer-Associated Diabetogenic Factor

Abstracts
2001 Digestive Disease Week

# 1793 A 2026 D Peptide as the Putative Pancreatic Cancer-Associated Diabetogenic Factor
Daniela Basso, Anna Valerio, Roberta Seraglia, Saverio Mazza, Eliana Greco, Carlo-Federico Zambon, Maria Grazia Piva, Nicoletta Gallo, Paola Fogar, Sergio Pedrazzoli, Antonio Tiengo, Mario Plebani, Padova, Italy

Aims of this study were to: 1) Verify whether culture media conditioned (CM) by different pancreatic cancer (PC) cell lines (CL) are able to alter glucose metabolism of isolated and perfused rat hepatocytes (IPRH); 2) Separate PC CM into fractions with different molecular weights (mw) and assess their metabolic effects on IPRH; 3) Analyze the molecular composition of control (NCM) and CM by mass spectrometry analysis (MALDI); 4) Verify whether any of these components is also detectable in the sera of control subjects (CS, n=10), pts with chronic pancreatitis (CP, n=9), or with pancreatic cancer (PC, n=14). The association with diabetes mellitus was also considered.

Methods: the following PC CL were used: MIA PaCa2, PSK, PANC-1, and CAPAN-1. The cells (about 100,000/75 cm2 flask) were cultured in appropriate low glucose cell culture media additioned with 10% FCS for 6 days; the culture media were then centrifuged and the supernatants collected (CM) for the experiments with IPRH; NCM was used as control. Two fractions for CM and NCM, one with a mw <30,000 D and another <10,000 D, were obtained after two steps filtration. IPRH, isolated from fasting male Wistar rats (130-250 g b.w.), were incubated with NCM or CM and glucose 20 mM. The IPRH suspension was sampled at time intervals for 2 hours. In the supernatants glucose and lactate were measured.

Results: In all experimental conditions, glucose concentration declined similarly over time, while lactate production was inhibited when the IPRH were incubated with the CM from all the PC cell lines. This inhibitory effect was reproduced also by CM with a mw <10,000 D. After MALDI analysis of this fraction, the following common peptides (pep)were identified in CM and NCM: 1667, 1795, 1952, 2065, 2144, 2193, 2272 and 2399D. A pep of 1874 D was expressed by all CM, another of 2026 D was expressed by 3/4 PC CL, and a 2726 D pep was expressed by 2/4 PC CL. Other low molecular weight pep were occasionally expressed by single PC cell lines. The pep with a mw of 2026, 2397 and 2726 D were found in patient's sera as follows: the 2026 D pep was identified in 1/10 CS, 7/14 PC, and 3/9 CP; the 2397 D pep in 5/10 CS, in 12/14 PC, and in 6/9 CP, the 2726 D pep in 0/10 CD, 11/14 PC, and 3/9 CP. Only the 2026 D pep was found to be associated with the presence of diabetes (Fisher's exact test:p<0.05).

Conclusions: The production of a diabetogenic factor is common to several PC cell lines. A component of 2026 D is significantly correlated with PC and diabetes, while another of 2726 D is tumor, but not diabetes associated.





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