Presence of CCK-A, B receptors and effect of gastrin and cholecystokinin on growth of pancreatobiliary cancer cell lines

Abstract

Aim: To investigate the effects of gastrin and cholecystokinin (CCK) and their specific antagonists on the growth of pancreatic and biliary tract cancer cell lines.

Methods: Five pancreatic and 6 biliary cancer cell lines with 2 conrtol cells were used in this study. Cell proliferation study was done using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test and direct cell count method. Reverse transcription-polymerase chain reaction (RT-PCR) and slot blot hybridization were performed to examine and quantify the expression of hormonal receptors in these cell lines.

Results: SNU-308 showed a growth stimulating effect by gastrin-17, as did SNU-478 by both gastrin-17 and CCK-8. The trophic effect of these two hormones was completely blocked by specific antagonists (L-365, 260 for gastrin and L-364, 718 for CCK). Other cell lines did not respond to gastrin or CCK. In RT-PCR, the presence of CCK-A receptor and CCK-B/gastrin receptor mRNA was detected in all biliary and pancreatic cancer cell lines. In slot blot hybridization, compared to the cell lines which did not respond to hormones, those that responded to hormones showed high expression of receptor mRNA.

Conclusion: Gastrin and CCK exert a trophic action on some of the biliary tract cancers.

Figure 1

Effects of gastrin and CCK on SNU-308 from gallbladder cancer (A), SNU-478 from ampulla of Vater cancer (B). ^aP<0.05 vs control.

Figure 2

Growth curves for SNU-308 (A), and SNU-478 (B, C) cells grown with either hormone or hormone combined with antagonist or antagonist, or medium only. G: gastrin-17, G*: antagonist for CCK-B/gastrin receptor (L-365,260), C: CCK-8, C*: antagonist for CCK-A receptor (L-364,718).

Figure 3

Amplification of CCK-A and CCK-B receptor mRNA in human biliary tract cancer cell lines (A) and pancreatic cancer cell lines (B) by RT-PCR.

Figure 4

Representative slot blot hybridization. (A) Data from SNU-308, 478, 1079, 410, Mia PaCa-2, and LoVo cell lines. (B, C) Densitometrically quantified slot blots.