cholecystokinin and Adenocarcinoma

Colorectal cancer is one of the leading causes of cancer-associated death in the United States and United Kingdom. In England and Wales, it is the second most common cancer in women and the third most common in men. Currently, treatment options for this debilitating disease are limited and surgical resection is the only curative treatment available. Despite rapid advances in surgery, as well as in adjuvant therapies such as radiotherapy and chemotherapy, there has been only a relatively modest improvement in mortality. The majority of colorectal cancers are epithelial-derived adenocarcinomas and arise from benign adenomas through the gain of mutations in key genes. Gastrin, an important polypeptide hormone, responsible for gastric acid secretion has been found to be involved in tumourigenesis in the gastrointestinal tract. When aberrantly expressed, the gastrin and gastrin/CCK-2 receptor genes can mediate powerful down stream events; the gastrin gene can impart anti-apoptotic properties while the gastrin/CCK-2 receptor can activate the transcription of a number of factors including ligands of the epidermal growth factor (EGF) receptor, the REG protein and matrix metalloproteinases (MMPs). In colonic tumourigenesis, gene expression of both gastrin and the gastrin/CCK-2 receptor is activated within epithelial cells at an early stage of the adenoma-carcinoma sequence. This review details the role played by gastrin in the adenoma-carcinoma sequence of colorectal carcinogenesis.

Topics: Adenocarcinoma; Cholecystokinin; Colorectal Neoplasms; Gastrins; Gene Expression Regulation, Neoplastic; Humans

The gut hormone cholecystokinin exerts various actions on the gastrointestinal tract, including the regulation of growth. The hormone has been reported to induce hypertrophy and hyperplasia of the pancreas and to enhance chemically-induced pancreatic carcinogenesis in animals. Stimulation of endogenous cholecystokinin secretion through the induction of deficiency of intraintestinal proteases and bile salts by trypsin-inhibiting nutrients, bile salt-binding drugs or surgical intervention is also capable of stimulating growth and tumour development in the rat. In man, factors suggested to increase the risk of pancreatic cancer, such as a high-fat and high-protein diet or gastrectomy, are known to stimulate plasma cholecystokinin secretion. Receptors for cholecystokinin have been demonstrated on human pancreatic adenocarcinomas, and cholecystokinin has been demonstrated to enhance the growth of xenografted pancreatic cancer and to inhibit growth of gastric and bile duct cancer. The recently developed cholecystokinin-receptor antagonists inhibit not only pancreatic growth but also pancreatic carcinogenesis in animals. These new drugs may be valuable new tools for inhibiting pancreatic cancer growth in humans.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Biliary Tract Neoplasms; Cholecystokinin; Gastrointestinal Neoplasms; Humans; Pancreatic Neoplasms

MK-329 is a nonpeptidal, highly specific cholecystokinin (CCK) receptor antagonist, with affinity for pancreatic and gallbladder CCK receptors similar to CCK itself. MK-329 and its progenitor, asperlicin, can inhibit the growth of CCK receptor-positive human pancreatic cancer in athymic mice. Based on these activities and the ability of MK-329 to transiently increase food intake and enhance morphine analgesia in murine models, we conducted an open trial of MK-329 in 18 patients with advanced pancreatic cancer in whom the CCK receptor status of the tumors was unknown. Tumor response, pain control, and nutritional parameters (hunger rating, caloric intake, body weight, and anthropometrics) were serially assessed. The results of the study failed to demonstrate any impact of MK-329 on tumor progression, pain, or nutrition. Toxicity was mild and limited to nausea, vomiting, diarrhea, and abdominal cramps, with 17 of 18 patients able to tolerate treatment. While a role for MK-329 in the management of patients with advanced pancreatic cancer cannot be supported by the results of this trial, additional studies of this agent in patients with known CCK receptor-positive tumors, at escalated doses, and possibly in conjunction with other growth antagonists, appear warranted.

Topics: Adenocarcinoma; Adult; Aged; Analgesia; Benzodiazepinones; Cholecystokinin; Devazepide; Female; Humans; Male; Middle Aged; Nutritional Physiological Phenomena; Pancreatic Neoplasms; Receptors, Cholecystokinin

Secretin and cholecystokinin (CCK) have trophic effects on the pancreas and may therefore have a place in the treatment of pancreatic cancer. The present study was performed to examine whether these hormones may cause harm in patients with pancreatic cancer receiving cytostatics. The cytostatics were 5-fluorouracil, adriamycin, and mitomycin C(FAM). Secretin plus Thr28Nle31CCK25-33, in doses stimulating pancreatic secretion to about 60% of maximal, were given as a continuous 6-day intravenous infusion just before (four patients) or immediately after (five patients) starting treatment with FAM. Five patients received FAM only. When considering symptoms, laboratory findings, abdominal CT scans, and survival, no evidence was found that secretin and CCK may cause serious or unpleasant side effects in patients with pancreatic cancer receiving cytostatics.

Topics: Adenocarcinoma; Aged; Alanine Transaminase; Alkaline Phosphatase; Amylases; Antineoplastic Combined Chemotherapy Protocols; Bilirubin; Cholecystokinin; Doxorubicin; Drug Evaluation; Female; Fluorouracil; Humans; Male; Middle Aged; Mitomycin; Mitomycins; Pancreatic Neoplasms; Peptide Fragments; Pilot Projects; Secretin

Gastrin, which induces gastric acid secretion, and a structurally similar hormone, cholecystokinin (CCK)-a potent acid inhibitor, may each play a role in gastric cancer. However, few studies have investigated this hypothesis in humans. We therefore investigated whether serum gastrin or CCK concentrations at baseline were associated with the incidence of gastric non-cardia adenocarcinomas (GNCA), oesophagogastric junctional adenocarcinomas (EGJA) or gastric carcinoid tumours over 24 years of follow-up in a study nested within the all-male Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study of Finnish smokers.. Totals of 283 incident GNCA, 96 EGJA and 10 gastric carcinoid cases, and 778 matched controls, were included in our analysis. Gastrin and CCK were measured using specific radioimmunoassays. Odds ratios (OR) and 95% confidence intervals (95% CI) were estimated by multivariable logistic regression with adjustment for all known or suspected confounding factors, including Helicobacter pylori seropositivity.. Those with high gastrin (Q4 vs Q1), had an increased risk of GNCA (fully adjusted OR: 1.92; 95% CI: 1.21, 3.05) and gastric carcinoids, though the small number of carcinoid cases meant the fully adjusted model was unstable (age-adjusted continuous model OR: 4.67; 95% CI: 2.67, 8.15). CCK was associated with risk of GNCA only for those in Q3 relative to Q1 (OR: 0.56; 95% CI: 0.33, 0.96), and no significant trend was observed.. Our data suggest that high serum concentrations of gastrin may be associated independently with an increased risk of gastric cancer; the role of CCK in cancer risk is less clear.

Topics: Adenocarcinoma; Case-Control Studies; Cholecystokinin; Esophageal Neoplasms; Finland; Follow-Up Studies; Gastrins; Helicobacter Infections; Helicobacter pylori; Humans; Incidence; Logistic Models; Male; Middle Aged; Multivariate Analysis; Prospective Studies; Risk Factors; Smokers; Stomach Neoplasms

Cholecystokinin (CCK) and gastrin stimulate growth of pancreatic cancer. Although down-regulation of gastrin inhibits growth of pancreatic cancer, the contribution of endogenous CCK to tumor growth is unknown. The purpose of this study was to evaluate the role of endogenous CCK on autocrine growth of pancreatic cancer. Pancreatic cancer cell lines were analyzed for CCK mRNA and peptide expression by real-time RT-PCR and radioimmunoassay, respectively. The effect of endogenous CCK on growth was evaluated by treating cancer cells with CCK neutralizing antibodies and by down-regulating CCK mRNA by RNAi. Wild-type pancreatic cancer cells expressed significantly lower CCK mRNA and peptide levels than gastrin. Neither treatment of pancreatic cancer cells with CCK antibodies nor the down-regulation of CCK mRNA and peptide by shRNAs altered growth in vitro or in vivo. Conversely, when gastrin mRNA expression was down-regulated, the same cells failed to produce tumors in spite of having sustained levels of endogenous CCK. Pancreatic cancer cells produce CCK and gastrin; however, the autocrine production of gastrin is more important for stimulating tumor growth.

Topics: Adenocarcinoma; Animals; Antibodies, Neutralizing; Autocrine Communication; Cell Line, Tumor; Cell Proliferation; Cholecystokinin; Gastrins; Gene Expression Regulation, Neoplastic; Humans; Male; Mice; Mice, Nude; Pancreatic Neoplasms; RNA, Messenger; RNA, Small Interfering; Tissue Distribution; Xenograft Model Antitumor Assays

To determine the immunoreactivity of cholecystokinin (CCK) during the development of the human fetal pancreas and pancreatic adenocarcinoma, given that, CCK positive cells were demonstrated either in its embryonic anlage or in pancreatic cancer. In order to obtain possible parallels in the expression pattern of neoplastic cells in adults (well--moderately--poorly differentiated), we investigated the pattern of CCK expression in the pancreatic tissue during the various stages of development and compared these with the proliferation of tissue assessed by proliferating cell nuclear antigen (PCNA) immunohistochemistry.. Tissue sections from 15 pancreatic fetal specimens, and equal number of ductal adenocarcinoma specimens, were assessed using immunohistochemical methods for CCK.. The density of positive cells in the primitive exocrine ductal walls and outgrowing buds was significantly higher than the relevant density in the neoplastic pancreatic tissue of mixed (ductal-endocrine) and pure ductal type (p1=0.004, p2 < 0.0005, p3 < 0.0005 and p4=0.023 respectively). The above values were estimated from 20th to 22nd weeks of gestation. There was no significant difference in the density of positive cells in the islet cell epithelium from 25-30 weeks, and the neoplastic tissue of mixed (p5=0.10) and pure ductal type (p6=0.15).. The immunostaining for CCK identifies a sub-group of pancreatic ductal adenocarcinomas with a neuroendocrine component (initially considered as pure ductal tumors), and mixed ductal-endocrine tumors. This pattern of expression in neoplasms recapitulates the normal pattern during the embryonal development of the organ, and may be important for the development of new therapeutic approaches with eventual clinical utility.

Topics: Adenocarcinoma; Aged; Cholecystokinin; Female; Gestational Age; Humans; Immunohistochemistry; Male; Middle Aged; Pancreas; Pancreatic Neoplasms

The cholecystokinin (CCK)/secretin pancreatic function tests to diagnose pancreatic exocrine insufficiency are time consuming and invasive. Our aim was to develop a rapid pancreatic function test performed during upper endoscopy that could discriminate between patients with normal from impaired exocrine pancreatic secretion.. We prospectively evaluated 412 patients for possible pancreatic diseases. During upper endoscopy, 1 CU/kg of secretin was given intravenously and duodenal juice (collected for 10 min) was assayed for concentrations of bicarbonate and lipolytic and trypsin activity. Final diagnosis was by histology, imaging, and a previously validated scoring system (for chronic pancreatitis). Of 412 patients, 117 patients had normal pancreas, 72 patients had chronic pancreatitis, and 116 patients had pancreatic adenocarcinoma. The remaining 107 patients had miscellaneous disease of the peripancreatic region. In 28 patients we also validated the secretin test with the standard CCK pancreatic function test.. There was no difference between bicarbonate or trypsin concentrations among the groups. Lipolytic concentration was significantly lower in chronic pancreatitis (115 +/- 18) and in pancreatic adenocarcinoma (87 +/- 10) compared with patients with normal pancreas (229 +/- 23; P < 0.03 and P < 0.0001, respectively). The overall accuracy of the endoscopic secretin test was 79%, with positive and negative predictive values of 73% and 85%, respectively. The concentration of lipolytic activity obtained by the endoscopic secretin test in 28 patients correlated moderately well (r = 0.41, P < 0.03) with lipolytic output obtained by the CCK pancreatic function test.. Lipolytic concentration in duodenal juice after intravenous secretin collected for 10 minutes during upper endoscopy was significantly lower in chronic pancreatitis and pancreatic adenocarcinoma compared with patients with normal pancreas, but was not accurate enough for routine clinical use.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Bicarbonates; Cholecystokinin; Chronic Disease; Diagnosis, Differential; Endoscopy, Gastrointestinal; Female; Humans; Lipase; Male; Middle Aged; Pancreatic Function Tests; Pancreatic Juice; Pancreatic Neoplasms; Pancreatitis; Secretin; Sensitivity and Specificity; Trypsin

Topics: Adenocarcinoma; Aged; Case-Control Studies; Cholecystokinin; Female; Gastrins; Humans; Male; Middle Aged; Pancreatic Neoplasms

It has been assumed that gastrin stimulates the growth of pancreatic cancer in an autocrine way through co-expression of gastrin and the cholecystokinin-B receptor (CCK-BR). However, pancreatic cancer cell lines established directly from patients have revealed a great heterogeneity in cell proliferation when exposed to CCK, gastrin and their receptor antagonists. The aim of this study was therefore to examine co-expression of CCK-A and CCK-B receptor (CCK-AR and CCK-BR), and gastrin mRNA as well as the secretion of CCK and gastrin peptides in these cell lines.. Fourteen cell lines were established from primary pancreatic cancers or their metastases. Total RNA was isolated from the cell lines and reverse-transcribed into single-stranded cDNA. A PCR technique based on Taq polymerase-antibody interaction and CCK-AR, CCK-BR and gastrin-specific primers, followed by Southern blot analysis, were the methods used. The incubation mediums were analysed for the presence of secreted CCK/proCCK and gastrin/progastrin peptides by specific radioimmunoassays (RIA).. By means of nested Reverse-Transcribed Polymerase Chain Reaction (nested RT-PCR), combined with Southem blot analysis of the PCR amplified products, CCK-AR and gastrin mRNA co-expression was detected in cell lines LPC-6p and LPC-10m, whereas CCK-BR and gastrin mRNA could be detected in cell lines LPC-8p and LPC-12m. A low level of secreted CCK peptides was detected in cell line LPC-6p, which also expressed CCK-AR mRNA. In no other cases were CCK or gastrin peptides detected in the cell culture mediums.. The lack of CCK-BR and gastrin mRNA co-expression, and not detectable levels of secreted CCK and gastrin in culture media, does not lend support to the hypothesis that concomitant gene-expression of CCK receptors and gastrin or CCK are essential to maintaining pancreatic cancer cell proliferation.

Topics: Adenocarcinoma; Adenocarcinoma, Mucinous; Aged; Biopsy; Blotting, Southern; Carcinoma, Ductal, Breast; Carcinoma, Papillary; Cell Division; Cholecystokinin; Gastrins; Gene Expression Regulation, Neoplastic; Humans; Middle Aged; Neoplasm Staging; Pancreatic Neoplasms; Radioimmunoassay; Receptors, Cholecystokinin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tumor Cells, Cultured

Alpha-amidated gastrin promotes the growth of nontransfected pancreatic cell lines expressing the gastrin/cholecystokinin (CCK)-B receptor. Gastrin/CCK-B and CCK-A receptors recently were demonstrated in human pancreatic adenocarcinomas, but to the authors' knowledge expression of their ligands to date have not been adequately investigated. As a prerequisite for making suggestions regarding local growth stimulation, the authors examined whether gastrin and the homologous CCK peptides as well as their specific receptors were expressed in consecutively collected solid human pancreatic adenocarcinomas.. Using a library of radioimmunoassays specific for different epitopes on proCCK, progastrin, their processing intermediates, and bioactive end products, CCK and gastrin gene expression was measured in extracts of solid human pancreatic adenocarcinomas (n = 19), resection margins (n = 15), and normal pancreatic tissue (n = 8). Moreover, CCK, CCK-A receptor, and gastrin/CCK-B receptor mRNA were measured by reverse transcriptase-polymerase chain reaction.. Amidated gastrins were synthetized in 14 of 19 carcinomas (median, 0.4 pmol/g; range, < 0.1-84.0 pmol/g) and in 12 of 15 resection margin samples (median, 0.3 pmol/g; range, < 0.1-6.1 pmol/g). In contrast, normal human pancreatic tissue expressed only traces of poorly processed progastrin. Gastrin/CCK-B receptor mRNA was present in all carcinomas, resection margins, and normal pancreatic tissue. CCK-A receptor mRNA was detected in most tumors, but neither the mature ligands (alpha-amidated and O-sulfated CCK peptides) nor their precursors were expressed in carcinoma and normal pancreatic tissue.. The results of the current study demonstrate that alpha-amidated gastrin peptides and their receptor invariably are coexpressed in pancreatic adenocarcinoma. Therefore these findings support the contention of a role for local gastrin regulatory mechanisms, but no CCK mechanisms, in pancreatic carcinoma.

Topics: Adenocarcinoma; Cholecystokinin; Humans; Neoplasm Proteins; Pancreas; Pancreatic Neoplasms; Receptor, Cholecystokinin A; Receptor, Cholecystokinin B; Receptors, Cholecystokinin; RNA, Messenger

For more than two decades, our research group has been studying the pancreatic actions of three groups of regulatory peptides: members of the cholecystokinin/gastrin family, bombesin-like peptides and somatostatin. Investigating these peptides, our work has focused on three particularly interesting aspects: peptidergic regulation of pancreatic enzyme secretion and growth in adult rats, peptidergic control of pancreatic enzyme secretion and growth during postnatal development in rats, and peptidergic regulation of proliferation and differential gene expression in pancreatic adenocarcinoma cells. Our data confirmed that the control of the exocrine function of the pancreas is complex, and that it involves peptides such as the cholecystokinin/gastrin-like peptides, bombesin-like peptides and somatostatin. In these investigations, it became evident that selective peptide receptor agonists, antagonists and monoclonal antibodies raised against peptides are useful tools to identify the role of these bioactive peptides in pancreatic exocrine secretion and cell proliferation.

Topics: Adenocarcinoma; Animals; Bombesin; Cell Division; Cholecystokinin; Gastrins; Gene Expression; Pancreas; Pancreatic Neoplasms; Rats; Somatostatin

Cholecystokinin (CCK) plays an important role in pancreatic carcinogenesis. While human CCK-A and -B receptors have been fully characterized, their relative roles in human pancreatic adenocarcinoma remain unclear. Thus, expression of CCK-A and -B receptors in normal human pancreas, pancreatic adenocarcinomas, and other human extrapancreatic tissues and malignancies was examined, using reverse transcription followed by the polymerase chain reaction (RT-PCR). mRNA isolated from 15 normal pancreas specimens, 22 pancreatic adenocarcinomas, and 58 extrapancreatic tissues and tumors was subjected to RT-PCR using primers specific for human CCK-A and -B receptors. Expression of CCK-B receptors was detected in all tissues arising from pancreas and in most extrapancreatic tissues and tumors. In contrast, CCK-A receptors exhibited a more selective pattern of expression in gall bladder, intestine, brain, ovary, spleen, and thymus. Of significance, CCK-A receptors were expressed selectively in all pancreatic adenocarcinomas, but not in any normal pancreas specimens. In situ hybridization, using receptor-specific riboprobes, localized CCK-A receptor expression to ductal cells, the presumed origin of most human pancreatic adenocarcinomas. Southern blot analysis revealed no evidence of CCK-A receptor gene amplification or rearrangement in pancreatic adenocarcinomas. Because of its selective expression, the CCK-A receptor may serve as selective biomarker for pancreatic adenocarcinoma.

Topics: Adenocarcinoma; Biomarkers, Tumor; Cholecystokinin; Humans; Pancreas; Pancreatic Neoplasms; Receptor, Cholecystokinin A; Receptor, Cholecystokinin B; Receptors, Cholecystokinin

Somatostatin inhibits proliferation of many solid tumors. The current study examines whether inhibition of the growth of pancreatic cancer by the somatostatin analog, octreotide, requires tumor expression of somatostatin receptors.. We studied five human pancreatic cancer cell lines, Capan-1, Capan-2, CAV, MIA PaCa-2, and Panc-1. Solid tumors were established in nude mice (n = 20/cell line) by flank injection of tumor cells. Subcutaneous octreotide (500 micrograms/kg/day) was administered by osmotic pumps to 10 of the animals in each group, and the other 10 received control infusions of saline solution. On day 36, the tumors were excised and weighed. Plasma levels of the putative trophic peptides cholecystokinin, epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1), and insulin were assessed by radioimmunoassay. Each of the five cell lines was assayed for the presence of cell surface somatostatin receptors by using whole cell competitive binding assays with 125I-somatostatin. Expression of the somatostatin receptor subtype-2 (SSR2) gene was determined with reverse transcriptase-polymerase chain reactions. Southern blot hybridization was used to assess the presence of the SSR2 gene.. Octreotide inhibited tumor growth in the MIA PaCa-2 group (512 +/- 75 mg control versus 285 +/- 71 mg treated; p < 0.05) but had no significant effect on tumor weight in the other four cell lines. Plasma levels of cholecystokinin, epidermal growth factor, insulin-like growth factor-1, and insulin were not altered by chronic octreotide infusion. Cell surface somatostatin receptors and SSR2 gene expression were detected only in the MIA PaCa-2 tumors. The gene for the SSR2 receptor was found in all five tumor lines.. Octreotide-mediated inhibition of pancreatic cancer growth is dependent on expression of somatostatin receptors. The expression of somatostatin receptors should be considered in the design and interpretation of clinical trials with somatostatin analogs for treatment of pancreatic cancer.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents, Hormonal; Base Sequence; Binding, Competitive; Blotting, Southern; Cell Division; Cholecystokinin; DNA, Neoplasm; Epidermal Growth Factor; Gene Expression Regulation, Neoplastic; Humans; Insulin; Insulin-Like Growth Factor I; Mice; Mice, Nude; Molecular Sequence Data; Octreotide; Pancreatic Neoplasms; Peptides; Polymerase Chain Reaction; Receptors, Somatostatin; Tumor Cells, Cultured

The growth of pancreatic cancers may be influenced by certain gut peptides. However, the alteration of gut peptide receptors in the progress of pancreatic carcinogenesis is largely unknown. With storage phosphor autoradiography, this study visualized and characterized receptors for cholecystokinin (CCK), somatostatin (SST), bombesin (BBS), secretin and vasoactive intestinal peptide (VIP) in pancreata of control hamsters (n = 7) and pancreatic preneoplastic lesions (n = 10) or adenocarcinomas (n = 10) of N-nitrosobis(2-oxopropyl)amine (BOP)-treated hamsters. The specific CCK-A and secretin receptors expressed in normal pancreata were markedly reduced in pancreatic preneoplastic lesions and absent in adenocarcinomas. In the development of pancreatic tumours, the subgroup of SST receptors did not change, but both the affinity and binding capacity declined. In comparison with the binding of VIP to normal pancreata, specific VIP binding was significantly lower in preneoplastic lesions and almost absent in pancreatic adenocarcinomas. No specific binding for BBS was detected in normal pancreas or (pre)neoplastic lesions of hamster pancreas. The reduction or absence of receptors for CCK, secretin, SST and VIP in hamster pancreas with the progress of carcinogenesis suggests that in BOP-treated hamsters, pancreatic adenocarcinomas have, to a large extent, lost the hormone-dependent characteristics of the original tissue.

Topics: Adenocarcinoma; Animals; Carcinogens; Cholecystokinin; Cricetinae; Mesocricetus; Nitrosamines; Pancreas; Pancreatic Neoplasms; Precancerous Conditions; Receptors, Cholecystokinin; Receptors, G-Protein-Coupled; Receptors, Gastrointestinal Hormone; Receptors, Somatostatin; Receptors, Vasoactive Intestinal Peptide; Secretin; Somatostatin; Vasoactive Intestinal Peptide

We compared morphological, biological and molecular biological patterns of a newly established, spontaneously immortalized pancreatic ductal cell line, TAKA-1, with a hamster pancreatic ductal adenocarcinoma cell line, PC-1. PC-1 cells grew in a monolayer on plastic tissue culture flasks, whereas TAKA-1 cells required type I collagen gel matrix to propagate. The growth rate and argyrophilic nuclear organizer region (Ag-NOR) counts were greater in PC-1 cells than in TAKA-1 cells. More TAKA-1 cells were in G0/G1 and less were in the S cell cycle phase than PC-1 cells. Karyotypically, the consistent change in TAKA-1 cells was an abnormal no. 3 chromosome, whereas additional chromosomal abnormalities were found in PC-1 cells. Ultrastructurally, TAKA-1 cells formed ductal structures and were composed of two types of cells, as in the normal hamster pancreatic ducts, whereas PC-1 cells were pleomorphic, showed evidence for loss of differentiation and contained intracytoplasmic lumens. Unlike the PC-1, TAKA-1 cells did not show a point mutation at codon 12 in the c-Ki-ras oncogene and did not grow in soft agar. Receptor binding assay showed specific epidermal growth factor binding to both cell lines, but secretin binding only to TAKA-1 cells. Both cells produced and released transforming growth factor-alpha in serum-free medium. Both cell lines expressed blood group A antigen, carbonic anhydrase, coexpressed cytokeratin and vimentin, and reacted with tomato and Phaseolus vulgaris leucoagglutinin (L-PHA) lectins. The results demonstrate that chromosomal abnormalities, cell cycle patterns, expression of cytokeratin 18, lectin bindings and the c-Ki-ras mutation are the features that distinguish the benign from the malignant pancreatic ductal cells in Syrian hamster.

Topics: Adenocarcinoma; Animals; Base Sequence; Cell Adhesion; Cell Cycle; Cell Division; Cell Line; Cholecystokinin; Cricetinae; Epidermal Growth Factor; Flow Cytometry; Gastrin-Releasing Peptide; Genes, ras; Immunohistochemistry; Karyotyping; Kinetics; Light; Mesocricetus; Microscopy; Microscopy, Electron; Molecular Sequence Data; Pancreas; Pancreatic Ducts; Pancreatic Neoplasms; Peptides; Point Mutation; Secretin; Tumor Cells, Cultured

We have studied the effects of acetylcholine (ACh), and other agents that modulate pancreatic bicarbonate secretion, on the anion permeability of a human ductal adenocarcinoma cell line (BxPC-3). Anion permeability was monitored using an 125I efflux assay. ACh (10 microM) markedly stimulated 125I efflux from BxPC-3 cells and this response was abolished by atropine (10 microM), indicating that it is mediated by muscarinic receptors. Using transport inhibitors and ionophores, we obtained data indicating that some of the ACh-induced 125I efflux results from the opening of K+ channels, which would hyperpolarise the cell and increase the electrical driving force for 125I exit. The remaining ACh-induced 125I efflux is not mediated by anion exchangers or by Na+/K+/2Cl- cotransporters, and is probably explained by activation of an anion channel in the BxPC-3 cell membrane. Ionomycin (0.5 microM) caused a small rise in 125I efflux, indicating that this process can be triggered by an increase in intracellular calcium concentration. ATP (100 microM), ADP (100 microM), bombesin (10 nM), and cholecystokinin (CCK) (10 nM) also stimulated 125I efflux, indicating that receptors for these agents are expressed on BxPC-3 cells. We speculate that bicarbonate secretion from the human pancreas could be modulated by ACh, ATP, bombesin, and CCK via a direct effect on the duct cell.

Topics: Acetylcholine; Adenine Nucleotides; Adenocarcinoma; Adenosine Triphosphate; Bombesin; Cholecystokinin; Humans; Iodine Radioisotopes; Ion Channels; Ionomycin; Pancreatic Neoplasms; Potassium Channels; Tumor Cells, Cultured

The Human pancreatic carcinoma cell line KP-1N and its clone KP-1NL which has a high rate of liver metastasis were established. Ki-ras DNA point mutation on the codon 12 was found. The growth of KP-1N was stimulated by a physiological range of concentration (10(-11)-10(-10) M) of cholecystokinin and the increase was inhibited by the addition of a cholecystokinin receptor antagonist (CR 1505). Daily injections of CR 1505 (35 mg/kg) diminished the number of tumor colonies in the liver that were formed after an intrasplenic injection of the highly liver metastatic KP-1NL cells. These results suggest that cholecystokinin antagonists may be useful as growth inhibitors for some pancreatic cancer.

Topics: Adenocarcinoma; Animals; Ceruletide; Cholecystokinin; Humans; In Vitro Techniques; Liver Neoplasms; Mice; Mice, Nude; Pancreatic Neoplasms; Proglumide; Tumor Cells, Cultured

Cholecystokinin (CCK) has been shown to increase cytosolic calcium and stimulate enzyme release from pancreatic acinar cells and a rat acinar cell line, AR42J. CCK is also trophic to normal pancreas and pancreatic cancer; however, the cellular mechanisms which regulate CCK-stimulated growth are unknown. The effect of CCK on intracellular calcium was evaluated in four human pancreatic cancer cell lines known to grow in response to CCK but not secrete enzymes (SW-1990, MIA PaCa-2, BXPC-3 and PANC-1) and a rat acinar cell line (AR42J) shown to secrete enzymes but not grow with CCK. By using single cell fluorescence microscopy in fura-2 loaded cells, intracellular calcium [Ca2+]i was measured. After obtaining baseline fluorescent cell images, synthetic CCK-octapeptide (CCK8) was added to the cells and images of cell fluorescence captured. [Ca2+]i of the rat acinar cells increased (603%) over the baseline within the first minute after the addition of CCK (4.10(-13) M to 4.10(-10) M) in 77% of cells tested. In contrast [Ca2+]i failed to significantly change in the human cancer cells treated with CCK. To further localize the defect in hormone signal transduction in cancer cells, cells were suspended in low calcium media and the plasma membranes were selectively permeabilized with digitonin. Media free calcium concentration was continuously monitored by fura-2 fluorescence. Addition of inositol 1,4,5-trisphosphate (IP3) resulted in a marked increase in medium calcium concentration indicating IP3 was capable of releasing calcium from intracellular stores in both the AR42J rat acinar cell line and in the human pancreas cancer cell lines. In conclusion, CCK does not increase cytosolic calcium in human pancreatic cancer cells in contrast to rat acinar cells although all contain IP3-sensitive intracellular Ca2+ pools. Our results suggest that growth promoting and secretory effects of CCK on pancreatic cells may occur via two independent signalling pathways.

Topics: Adenocarcinoma; Calcium; Cholecystokinin; Cytosol; Fura-2; Humans; Image Processing, Computer-Assisted; Inositol 1,4,5-Trisphosphate; Pancreas; Pancreatic Neoplasms; Signal Transduction; Tumor Cells, Cultured

The growth responses of six human pancreatic cancer cell lines (SW-1990, PANC-1, MIA PaCa-2, BxPC-3, RWP-2 and CAPAN-2) to cholecystokinin (CCK) were evaluated in serum-free medium (SFM). In each experiment cells were initially plated in media containing fetal calf serum (FCS) grown for 48-72 h, and then washed with saline. Cells were incubated for an additional 72 to 96 h in medium devoid of FCS in the absence (control) or presence of synthetic CCK analogue (Thr4,Nle7)CCK9 (10(-13) to 10(-9) M), or CCK8 (10(-12) to 10(-9) M), or CCK39 (10(-12) to 10(-9) M). Viable cell counts were performed with a hemocytometer. Growth of each cell line was stimulated in the presence of CCK in serum-free medium, although the magnitude of responses differed. The concentrations of (Thr4,Nle7)CCK9 which stimulated the greatest increase in cell counts as compared to controls for each cell line were: SW-1990, 39% (10(-12) M, P less than 0.05); PANC-1, 45% (10(-9) M, P less than 0.005); MIA PaCa-2, 42% (10(-12) M, P less than 0.005); BxPC-3, 32% (10(-13) M, P less than 0.05); RWP-2, 37% (10(-11) M, P less than 0.005). Maximal response to CCK8 occurred at the 10(-9) M dose for each cell line: MIA PaCa-2, 40% (P less than 0.025); PANC-1, 85% (P less than 0.001); RWP-2, 68% (P less than 0.001) and CAPAN-2, 52% (P less than 0.001). The maximal increase in cell count with CCK39 ranged from 44-74% and occurred with either 10(-11) or 10(-10) M. CCK8 in SFM also stimulated cell growth as well as or better than FCS alone in three out of four pancreatic cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenocarcinoma; Animals; Cell Count; Cell Transformation, Neoplastic; Cholecystokinin; Mice; Mice, Nude; Pancreatic Neoplasms; Tumor Cells, Cultured

Flow cytometry and immunohistochemical analyses of the human gastric adenocarcinoma cell line MKN45G identified an intracellular peptide recognized by an anti-gastrin-17 (G17) antiserum but not by an anti-cholecystokinin-specific antiserum. Staining was not associated with the parental line MKN45, of which MKN45G is a clonal variant. The MKN45G cell line had elevated in vitro growth in serum-free medium in which the proliferation of MKN45G cells but not MKN45 cells was reduced to 58% of the control value by treatment with a rabbit anti-G17 antiserum. This inhibition of proliferation was reversed by preabsorbing the antiserum with excess G17. Disaggregated primary human gastric and colorectal tumors were screened for gastrin immunoreactivity by flow cytometry, and 6 of 28 colorectal and 8 of 22 gastric tumors had greater than 20% positively staining cells.

Topics: Adenocarcinoma; Animals; Antibodies; Carcinoembryonic Antigen; Cell Division; Cholecystokinin; Colorectal Neoplasms; Flow Cytometry; Gastrins; Gastrointestinal Neoplasms; Humans; Immunohistochemistry; Intracellular Fluid; Rabbits; Stomach Neoplasms; Tumor Cells, Cultured

The effect of a synthetic analogue of CCK (Thr4,Nle7CCK-9) on growth of SW-1990 human pancreatic cancer was examined in two experimental models. Nude mice bearing SW-1990 pancreatic cancer xenografts were injected with CCK (5, 15, or 25 micrograms/kg) or vehicle twice daily for 20 days. Animals were then sacrificed and tumor volume, weight, protein, and deoxyribonucleic acid (DNA) content were evaluated. SW-1990 cells were grown in vitro and the effects of CCK, secretin, vasoactive intestinal peptide (VIP), and proglumide (a CCK-receptor antagonist) on cell number and DNA synthesis were determined. The highest dose of CCK, 25 micrograms/kg, significantly increased tumor weight, protein content, and DNA content (P less than 0.005). In vitro, CCK caused significant increases in cell counts of up to 47% at six days and 66% at 12 days compared to control. Graded concentrations of CCK had a biphasic effect on DNA synthesis with significant increases of up to 65% (P less than 0.005). CCK-induced cell proliferation was inhibited by proglumide. Secretin slightly increased cancer cell growth, although not as potently as CCK, VIP or secretin in combination with CCK did not show potentiation. These results indicate that growth of some human pancreatic cancers may be influenced by gastrointestinal peptides, of which CCK is the most potent.

Topics: Adenocarcinoma; Animals; Cholecystokinin; DNA, Neoplasm; Dose-Response Relationship, Drug; Humans; Male; Mice; Mice, Nude; Organ Size; Pancreas; Pancreatic Neoplasms; Proglumide; Secretin; Tumor Cells, Cultured; Vasoactive Intestinal Peptide

This study deals with the effect of four types of COOH-terminal cholecystokinin (CCK) fragments on the growth of xenotransplantable human gastric cancer (SC-6-JCK, a poorly differentiated adenocarcinoma) whose growth has been promoted by pentagastrin. The growth of the tumor was inhibited using daily s.c. injections of CCK-octapeptide (CCK-8) and glutaryl-CCK-8 at a dose of 500 micrograms/kg body weight. After 30 days of treatment with CCK-8 or glutaryl-CCK-8, a significant decrease was observed in the tumor weight (P less than 0.05) and the tumor size P less than 0.01) in comparison with those of the control. But treatment with CCK-12 and pyroglutamyl-CCK-8 did not produce inhibition of tumor growth. Furthermore the correlation between the effect of CCK-8 on the normal rise in tumor cyclic adenosine 3':5'-monophosphate (cAMP) levels caused by pentagastrin injection and tumor growth was studied. The increase of cAMP by a single i.p. injection of pentagastrin at a dose of 20 micrograms/mouse was significantly inhibited by pretreatment with CCK-8 at concentrations equimolar to pentagastrin (P less than 0.05), while cAMP in the tumor was slightly elevated by a single i.p. injection of CCK-8 alone. Also in the in vitro study, CCK-8 inhibited the increase of cAMP and the activation of cAMP-dependent protein kinase which was stimulated by pentagastrin. These results suggest that proliferation of gastrin-dependent human gastric cancers may be suppressed by CCK in competition with gastrin.

Topics: Adenocarcinoma; Animals; Cholecystokinin; Cyclic AMP; Humans; Mice; Neoplasm Transplantation; Pentagastrin; Protein Kinases; Sincalide; Stomach Neoplasms

Topics: Adenocarcinoma; Animals; Carcinogens; Cholecystokinin; Cricetinae; Disease Models, Animal; Neoplasm Metastasis; Nitrosamines; Pancreas; Pancreatic Juice; Pancreatic Neoplasms

The role of the pancreaticotrophic hormone cholecystokinin (CCK) in modifying the pancreatic response to carcinogen has been examined in the hamster-nitrosamine pancreatic cancer model. Exogenous CCK, 30 IDU kg-1, stimulated a maximal pancreatic secretory response when given intravenously and caused hypertrophy and hyperplasia of the pancreas when given subcutaneously over a period of 6 weeks (pancreatic wet weight, mg per 100 g body weight, controls 295.6 +/- 61; CCK treated 466.4 +/- 77, P less than 0.001). When the same dose of CCK was given to animals receiving N-nitrosobis (2-oxopropyl)amine (BOP; 5 mg kg-1 weekly) there was a reduction in latency period and increase in induction rate of tumour development (CCK + BOP vs. BOP alone, 12 animals with tumours vs. 2 at 15 weeks; P less than 0.02). These effects are consistent with CCK acting as a co-carcinogen or promoter of pancreatic carcinogenesis in this model.

Topics: Adenocarcinoma; Animals; Cholecystokinin; Cocarcinogenesis; Cricetinae; Male; Mesocricetus; Nitrosamines; Pancreas; Pancreatic Juice; Pancreatic Neoplasms; Precancerous Conditions

A primary endometrial adenocarcinoma is reported that showed abundant foci suggestive of pathologically differentiated intestinal epithelium. The tumor epithelium was composed of four main cell types. Columnar cells resembled absorptive intestinal cells and displayed glycocalyceal carcinoembryonic antigen immunostaining; mucin-producing cells and a few Paneth-like lysozyme-rich cells were irregularly distributed; a massive quantity of argyrophil cells including a few amphicrine (muco-argyrophil) ones, were detected by Grimelius-Alcian blue method. Immunocytochemical evidence was obtained for the storage of serotonin, somatostatin and gastrin/cholecystokinin in some of the endocrine cells. These findings suggest that the tumor arose from a pluripotential stem cell of the glandular epithelium.

Topics: Adenocarcinoma; Aged; Cholecystokinin; Female; Gastrins; Histocytochemistry; Humans; Immunochemistry; Somatostatin; Uterine Neoplasms

A primary adenocarcinoma of the nasal cavity with light microscopic, electron microscopic, and immunocytochemical features of an enteric-type carcinoma is presented. The carcinoma contained a variety of dense-core granules similar to those seen in enterochromaffin cells of different functional types. Some granules demonstrated an immunoreactivity with serotonin, cholecystokinin, gastrin, somatostatin and leu-enkephalin antibodies. It is suggested that the endocrine cells in the neoplasm belong to the non-neuroectodermal paraneurone system.

Topics: Adenocarcinoma; Aged; Cholecystokinin; Chromaffin System; Enterochromaffin Cells; Humans; Immunoenzyme Techniques; Male; Microscopy, Electron; Nasal Cavity; Nose Neoplasms; Serotonin; Vasoactive Intestinal Peptide

Topics: Acute Disease; Adenocarcinoma; Carcinoma; Cholangiopancreatography, Endoscopic Retrograde; Cholecystokinin; Chronic Disease; Humans; Insulinoma; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Secretin; Time Factors; Zollinger-Ellison Syndrome

In animal studies, DL-[carboxyl-14C]tryptophan [DL-Try(C-14)] showed a high specificity for the pancreas, which suggested the potential of DL-[carboxyl-11C]tryptophan [DL-Try(C-11)] for clinical pancreatic inaging. The blood clearance and tissue uptake of the amino acid were very rapid, and no carrier effect was observed through a dose of 5 mg/kg. None of three transplanted hamster pancreatic adenocarcinomas that we studied showed a selective uptake of DL-Try(C-14) by the tumor, and none of the three enzymatic regimens investigated gave significant enhancement of the pancreatic specificity. Commercial L-Try(C-14) gave slightly better pancreatic specificity than the analogous racemic compound but without enough improvement to warrant attempts at optical resolution. DL-Try(C-11) was synthesized in amounts up to 325 mCi using a rapid, high-temperature, high-pressure modification of the Bücherer-Strecker amino acid synthesis. Yields ranged from 30--60%, and a total of 40 min was required for synthesis and chromatographic purification. DL-Try(C-11) thus appears to have significant potential as a clinical pancreas-imaging agent, particularly when used in conjunction with positron computerized transaxial tomography.

Topics: Adenocarcinoma; Animals; Carbon Radioisotopes; Cholecystokinin; Cricetinae; Dogs; Female; Isotope Labeling; Male; Mesocricetus; Neoplasm Transplantation; Neoplasms, Experimental; Pancreas; Pancreatic Neoplasms; Rabbits; Radionuclide Imaging; Rats; Secretin; Stimulation, Chemical; Tissue Distribution; Transplantation, Homologous; Tryptophan