trypsinogen and Pancreatic-Neoplasms

Topics: Abdominal Pain; Adult; Age of Onset; Calculi; Child; Developing Countries; Diabetes Mellitus; Humans; Malnutrition; Manihot; Mutation; Oxidative Stress; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis, Chronic; Steatorrhea; Tropical Climate; Trypsin; Trypsinogen

Hereditary chronic pancreatitis (HCP) is a very rare form of early-onset chronic pancreatitis. Apart from young age at diagnosis and a slower progression, the clinical course, morphological features and laboratory findings of HCP do not differ from those of patients with alcoholic chronic pancreatitis. Diagnostic criteria and treatment of HCP also resemble those of chronic pancreatitis of other causes. The clinical presentation is highly variable and includes chronic abdominal pain, impairment of endocrine and exocrine pancreatic function, nausea and vomiting, maldigestion, diabetes, pseudocysts, bile-duct and duodenal obstruction, and rarely pancreatic cancer. Fortunately, the disease is mild in most patients. Mutations in the PRSS1 gene, encoding cationic trypsinogen, play a causative role in chronic pancreatitis. It has been shown that the PRSS1 mutations increase autocatalytic conversion of trypsinogen to active trypsin, and thus probably cause premature, intrapancreatic trypsinogen activation, disturbing the intrapancreatic balance of proteases and their inhibitors. Other genes--such as the anionic trypsinogen (PRSS2), the serine protease inhibitor Kazal type 1 (SPINK1), and the cystic fibrosis transmembrane conductance regulator (CFTR)--have also been found to be associated with chronic pancreatitis (idiopathic and hereditary). Genetic testing should only be performed in carefully selected patients by direct DNA sequencing, and antenatal diagnosis should not be encouraged. Treatment focuses on enzyme and nutritional supplementation, pain management, pancreatic diabetes, and local organ complications such as pseudocysts and bile-duct or duodenal obstruction. The disease course and prognosis of patients with HCP is unpredictable. The risk of pancreatic cancer is elevated. Therefore, HCP patients should strongly avoid environmental risk factors for pancreatic cancer.

Topics: Animals; Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Diagnosis, Differential; Genetic Counseling; Genetic Predisposition to Disease; Humans; Models, Animal; Mutation; Pancreatic Neoplasms; Pancreatitis, Chronic; Prenatal Diagnosis; Prognosis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Hereditary chronic pancreatitis (HCP) is a very rare form of early onset chronic pancreatitis. With the exception of the young age at diagnosis and a slower progression, the clinical course, morphological features and laboratory findings of HCP do not differ from those of patients with alcoholic chronic pancreatitis. As well, diagnostic criteria and treatment of HCP resemble that of chronic pancreatitis of other causes. The clinical presentation is highly variable and includes chronic abdominal pain, impairment of endocrine and exocrine pancreatic function, nausea and vomiting, maldigestion, diabetes, pseudocysts, bile duct and duodenal obstruction, and rarely pancreatic cancer. Fortunately, most patients have a mild disease. Mutations in the PRSS1 gene, encoding cationic trypsinogen, play a causative role in chronic pancreatitis. It has been shown that the PRSS1 mutations increase autocatalytic conversion of trypsinogen to active trypsin, and thus probably cause premature, intrapancreatic trypsinogen activation disturbing the intrapancreatic balance of proteases and their inhibitors. Other genes, such as the anionic trypsinogen (PRSS2), the serine protease inhibitor, Kazal type 1 (SPINK1) and the cystic fibrosis transmembrane conductance regulator (CFTR) have been found to be associated with chronic pancreatitis (idiopathic and hereditary) as well. Genetic testing should only be performed in carefully selected patients by direct DNA sequencing and antenatal diagnosis should not be encouraged. Treatment focuses on enzyme and nutritional supplementation, pain management, pancreatic diabetes, and local organ complications, such as pseudocysts, bile duct or duodenal obstruction. The disease course and prognosis of patients with HCP is unpredictable. Pancreatic cancer risk is elevated. Therefore, HCP patients should strongly avoid environmental risk factors for pancreatic cancer.

Topics: Adult; Animals; Carrier Proteins; Child; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Databases, Genetic; Diagnosis, Differential; Disease Models, Animal; Genetic Counseling; Genetic Predisposition to Disease; Genetic Testing; Humans; Mice; Mutation; Pancreatic Neoplasms; Pancreaticojejunostomy; Pancreatitis, Chronic; Prognosis; Rats; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Pancreatic Neoplasms; Pancreatitis, Alcoholic; Pancreatitis, Chronic; Point Mutation; Risk Factors; Smoking; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Trypsinogens and PSTI/TATI/SPINK1 are expressed, usually together, at high levels by the pancreas but also by many other normal and malignant tissues. The present review describes studies on the expression and putative functions of trypsinogens and PSTI/TATI/SPINK1 in the human body. The clinical aspects are discussed, including the correlations between expression of trypsinogens and PSTI/TATI/SPINK1 in tissues, serum, and urine of patients with pancreatitis or cancer and clinicopathological characteristics, i.e., the roles of trypsinogens and PSTI/TATI/SPINK1 in spontaneous and hereditary pancreatitis, tumor progression, and prognosis.

Topics: Biomarkers, Tumor; Humans; Neoplasms; Pancreas; Pancreatic Neoplasms; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

There was some recent progress in the understanding of genetic risk factors in chronic pancreatitis. Due to this progress some of the traditional views of the subject will change. Today, genetic risk factors are attributed a much more important role that in the past. The frequency and strength of mutations were higher than expected. Strong variants were the rare autosomal-dominant mutations N29I and R122H of PRSS1 (cationic trypsinogen) and homozygous N34S of SPINK1 (pancreatic secretory trypsin inhibitor). Other mutations (heterozygous N34S, CFTR) were of lower relevance but still mediate a higher risk than alcohol consumption. The course of genetically determined pancreatitis is rather mild. In the long term pancreas cancer was found in some patients but apart from non-smoking no adequate prophylactic strategy is available up to now.

Topics: Carrier Proteins; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Diabetes Mellitus; Disease Progression; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Cytogenetic Analysis; Humans; Intercellular Signaling Peptides and Proteins; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The importance of pretest information, using an accredited DNA laboratory and interpreting the genotype on behalf of the patient and their physicians is emphasized. Care with predictive testing and the strong encouragement to involve a specialist genetic counseling service is made. A similar approach to genetic testing should be used when children are involved. Because of the incomplete pickup of PRSS1 mutations, particularly of a limited mutation panel of R122H and N291 (perhaps with A16V), a diagnosis of HP cannot be ruled out by molecular genetic testing alone. The A16V mutation has a reduced penetrance, and its contribution to pancreatitis remains unclear. The advice to patients with genetic forms of pancreatitis is a strong encouragement to avoid smoking, to avoid alcohol, and to remain in contact with clinical and research groups for their follow-up and screening trials for early pancreatic cancer. The remaining issues are of how wide to cast the net of investigation in patients with unexplained pancreatitis, particularly looking for mutations in the CFTR and lower penetrance genes such as PSTI/SPINK1.

Topics: Algorithms; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Genetic Counseling; Genetic Testing; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Pregnancy; Prenatal Diagnosis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The classical feature of hereditary pancreatitis (HP) is characterized by recurrent episodes of acute pancreatitis or a priori chronic pancreatitis in several members of one family. In 1996, the identification of the first HP-associated mutation in the cationic trypsinogen gene provided a breakthrough in our understanding of the pathogenesis of chronic pancreatitis. In the following years, several different mutations in the same gene have been found in a large number of investigated families. Most intriguing, HP patients have a more than 50-fold increased risk of pancreatic ductal cancer in comparison with expected pancreatic cancers in the general population. Variants of the major intrapancreatic trypsin antagonist SPINK1 have implications for more common forms of chronic pancreatitis. Research has focussed on the SPINK1-N34S-mutation, which is closely associated with tropical, alcoholic, or "idiopathic" chronic pancreatitis. Chronic pancreatitis represents a variable part of the cystic fibrosis syndrome, which is caused by mutations in the gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR). Several groups have reported an increased prevalence of CFTR mutations in patients with chronic pancreatitis of different etiology. In this review, we summarize interesting clinical and biochemical features of genetic variants in these genes which are associated with chronic pancreatitis.

Topics: Carrier Proteins; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Poland; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Hereditary pancreatitis (HP) is a disease which has been discovered quite recently. The inheritance is autosomally dominant with 80% penetrance. It gives the same symptoms as acute pancreatitis in early childhood and ends up with chronic pancreatitis. In 60% of the patients, a mutation in the trypsinogen gene can be demonstrated. The remaining 40% of the HP patients are diagnosed on the basis of clinical criteria. The acute and the chronic pancreatitis are treated as usual. It is important to recognize the disease because patients with HP have a 50 times increased risk of developing pancreatic cancer. At the age of 70, 40% have developed pancreatic cancer. This risk doubles for cigarette smokers. Screening programmes for HP in order to prevent pancreatic cancer are, however, expensive and troublesome.

Topics: Acute Disease; Adult; Aged; Child; Chronic Disease; Genetic Predisposition to Disease; Genetic Testing; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsinogen

Acute pancreatitis is a disorder that has numerous causes and an obscure pathogenesis. Bile duct stones and alcohol abuse together account for about 80% of acute pancreatitis. Most episodes of biliary pancreatitis are associated with transient impaction of the stone in the ampulla (that causes obstruction of the pancreatic duct, with ductal hypertension) or passage of the stone though and into the duodenum. Other causes of acute pancreatitis are various toxins, drugs, other obstructive causes (such as malignancy or fibrotic sphincter of Oddi), metabolic abnormalities, trauma, ischemia, infection, autoimmune diseases, etc. In 10% of cases of acute pancreatitis, no underlying cause can be identified; this is idiopathic pancreatitis. Occult biliary microlithiasis may be the cause of two thirds of the cases of "idiopathic" acute pancreatitis. Intra-acinar activation of trypsinogen plays a central role in the pathogenesis of acute pancreatitis, resulting in subsequent activation of other proteases causing the subsequent cell damage. Ischemia/reperfusion injury is increasingly recognized as a common and important mechanism in the pathogenesis of acute pancreatitis and especially in the progression from mild edematous to severe necrotizing form. Increased intracellular calcium concentration also mediates acinar cell damage. Oxygen-derived free radicals and many cytokines (e.g., interleukin [IL]-1, IL-6, IL-8, tumor necrosis factor-alpha, platelet activating factor) are considered to be principal mediators in the transformation of acute pancreatitis from a local inflammatory process into a multiorgan illness.

Topics: Acute Disease; Autoimmune Diseases; Child; Cholelithiasis; Female; Humans; Hypercalcemia; Male; Pancreas; Pancreatic Neoplasms; Pancreatitis; Pancreatitis, Alcoholic; Pregnancy; Reperfusion Injury; Trypsinogen

Mutations of the cationic trypsinogen gene have been detected in hereditary pancreatitis. This article reviews current understanding of their function and clinical significance.. An unrestricted Medline search was conducted using the key words hereditary pancreatitis and 'cationic trypsinogen . Additional material was obtained from references cited in original papers and recently published abstracts of meetings.. Cationic trypsinogen mutations have been identified in most, but not all, families with hereditary pancreatitis. This confirms existing evidence that premature trypsinogen activation plays a central role in the pathogenesis of human pancreatitis. Patients currently clinically defined as having hereditary pancreatitis should be screened for the presence of cationic trypsinogen mutations. A subgroup of patients with non-hereditary pancreatitis may also benefit from being screened for these mutations. Patients with hereditary pancreatitis should be entered into prospective, multicentre trials investigating secondary screening for pancreatic cancer. Gene therapy for hereditary pancreatitis is beyond current technological capability but remains a future therapeutic prospect for this often debilitating condition.

Topics: Acute Disease; Cations; Chronic Disease; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsinogen

Hereditary pancreatitis is a rare condition characterized by acute and chronic pancreatitis transmitted in an autosomal dominant fashion. There also is an epidemiologic link to pancreatic cancer in some affected families. Failure of a secondary brake mechanism responsible for inactivation of prematurely activated cationic trypsin in acinar cells seems to be the fundamental defect in type I hereditary pancreatitis (R117H cationic trypsin), and also may explain the pathogenesis of type II hereditary pancreatitis (N211 cationic trypsin). The diagnosis is made based on clinical history and, in certain cases, by molecular diagnostic testing for these gene defects. Medical management of acute and chronic hereditary pancreatitis currently does not differ from that of nonhereditary AP. As in nonhereditary pancreatitis, the surgical approach must be tailored to the individual problem, with an understanding that disease restricted to the head of the gland is atypical and that residual acinar tissue continues to drive the disease state. Although diagnosis and management of pancreatic adenocarcinoma are similar in this cohort, the increased age-accumulated risk suggests that thoughtful screening protocols eventually may be clinically and cost-effective.

Topics: Adenocarcinoma; Chromosomes, Human, Pair 7; Female; Humans; Male; Pancreatic Neoplasms; Pancreatitis; Point Mutation; Trypsinogen

Hereditary pancreatitis is an unusual form of acute and chronic pancreatitis that is usually associated with two specific mutations in the cationic trypsinogen gene. The extensive information available on the biochemistry, cell biology, and molecular biology of cationic trypsinogen and its gene provides the groundwork for development of a variety of therapeutic strategies, including gene therapy. Several features of this disease, however, make gene therapy unlikely in the near future. Further research using new models, including transgenic animals, is required before breakthroughs in therapy can be expected.

Topics: Amino Acid Sequence; Animals; Chymotrypsinogen; Genetic Therapy; Humans; Molecular Sequence Data; Mutation; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

The recent discovery that mutations in the trypsinogen gene are responsible for acute and chronic pancreatitis, and that patients with hereditary pancreatitis are at great risk for pancreatic cancer, has opened the door to understanding many aspects of pancreatic disease. This review focuses on the clinical presentation of hereditary pancreatitis, the mechanism of disease, and implications of this disease on understanding acute and chronic pancreatitis.

Topics: Acute Disease; Amino Acid Substitution; Chronic Disease; Genetic Predisposition to Disease; Humans; Mutation, Missense; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Hereditary pancreatitis is an unusual form of acute and chronic pancreatitis with a familial predisposition. Recently, the gene mutations causing most cases of hereditary pancreatitis have been identified in the cationic trypsinogen gene. The known mutations are trypsinogen R117H and N211. These may predispose to acute pancreatitis by eliminating one of the fail-safe mechanisms used by the pancreas to eliminate prematurely activated trypsin. Accumulation of active trypsin mutants are hypothesized to initiate a digestive enzyme activation cascade in the pancreatic acinar cells leading to autodigestion, an intense inflammatory response, and acute pancreatitis. The observation that these patients also develop typical chronic pancreatitis and may later develop pancreatic cancer provides strong evidence that these conditions are linked. Knowledge of the pathophysiological conditions leading to acute and chronic pancreatitis and the development of a transgenic mouse expressing the mutant human trypsinogen genes will provide directions and tools necessary for the effective treatment or prevention of this human disease.

Topics: Animals; Chronic Disease; Disease Models, Animal; Genetic Diseases, Inborn; Humans; Mice; Pancreatic Neoplasms; Pancreatitis; Prognosis; Sensitivity and Specificity; Trypsinogen

Important advances in the understanding of pancreatic diseases have taken place through the application of molecular methods in the study of the inherited form of pancreatitis and pancreas cancer. Mutations of the cationic trypsinogen gene have been found to be causative for hereditary pancreatitis with important implications for the molecular pathogenesis of acute and chronic pancreatitis. A variety of cancer syndromes involving the P16 and BRCA2 genes, for example, also lead to pancreatic cancer, but the gene responsible for familial pancreatic cancer has not been identified so far. The establishment of a European Registry of Hereditary Pancreatitis and Pancreatic Cancer (EUROPAC) will facilitate future developments.

Topics: Family Health; Humans; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Increased serum concentrations of trypsin immunoreactivity occur in patients with biliary tract cancer. To characterize this trypsin, we developed a sensitive time-resolved immunofluorometric assay for trypsin-1 complexed with alpha(1)-antitrypsin (AAT) and studied the concentrations of this complex in sera from healthy individuals (n = 130) and patients with benign biliary disease (n = 32), biliary tract cancer (n = 17), pancreatic cancer (n = 27), and hepatocellular cancer (n = 12).. We used a trypsin-1-specific monoclonal antibody on the solid phase and a europium-labeled polyclonal antibody to AAT as tracer. The detection limit was 0.42 microgram/L. The validity of the trypsin-1-AAT test for detection of biliary tract cancer was compared with trypsin-2-AAT and CA19-9.. Increased concentrations of trypsin-1-AAT (>33 microgram/L) were found in 76% of patients with biliary tract cancer, and the concentrations were significantly higher than in those with benign biliary disease (P <0. 0001). The median concentration of trypsin-1-AAT in serum from patients with biliary tract cancer was 3.7-fold higher than in healthy controls, 2.6-fold higher than in patients with benign biliary tract disease, 1.7-fold higher than in patients with pancreatic cancer, and 2.0-fold higher than in patients with hepatocellular cancer.. Of the markers studied, trypsin-1-AAT had the largest area (0.83) under the receiver operating curve in differentiating biliary tract cancer from benign biliary tract disease. Our results suggest that trypsin-1-AAT is a new potential marker for biliary tract cancer.

Topics: alpha 1-Antitrypsin; Biliary Tract Diseases; Biliary Tract Neoplasms; Biomarkers, Tumor; CA-19-9 Antigen; Fluoroimmunoassay; Humans; Isoenzymes; Pancreatic Neoplasms; Reproducibility of Results; Sensitivity and Specificity; Trypsin; Trypsinogen

Cancer stem cells (CSCs) subpopulation within the tumour is responsible for metastasis and cancer relapse. Here we investigate in vitro and in vivo the effects of a pancreatic (pro)enzyme mixture composed of Chymotrypsinogen and Trypsinogen (PRP) on CSCs derived from a human pancreatic cell line, BxPC3. Exposure of pancreatic CSCs spheres to PRP resulted in a significant decrease of ALDEFLUOR and specific pancreatic CSC markers (CD 326, CD 44 and CxCR4) signal tested by flow cytometry, further CSCs markers expression was also analyzed by western and immunofluorescence assays. PRP also inhibits primary and secondary sphere formation. Three RT

Topics: Animals; Cell Line, Tumor; Chymotrypsinogen; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Humans; MAP Kinase Signaling System; Mice; Mice, Nude; Neoplastic Stem Cells; Pancreatic Neoplasms; Trypsinogen; Xenograft Model Antitumor Assays

Pancreatic ductal adenocarcinoma (PDAC) is a very aggressive tumor with a five-year survival of less than 6%. Chronic pancreatitis (CP), an inflammatory process in of the pancreas, is a strong risk factor for PDAC. Several genetic polymorphisms have been discovered as susceptibility loci for both CP and PDAC. Since CP and PDAC share a consistent number of epidemiologic risk factors, the aim of this study was to investigate whether specific CP risk loci also contribute to PDAC susceptibility. We selected five common SNPs (rs11988997, rs379742, rs10273639, rs2995271 and rs12688220) that were identified as susceptibility markers for CP and analyzed them in 2,914 PDAC cases, 356 CP cases and 5,596 controls retrospectively collected in the context of the international PANDoRA consortium. We found a weak association between the minor allele of the PRSS1-PRSS2-rs10273639 and an increased risk of developing PDAC (OR

Topics: Adenocarcinoma; Adult; Aged; Biomarkers, Tumor; Carcinoma, Pancreatic Ductal; Case-Control Studies; Female; Follow-Up Studies; Humans; Male; Middle Aged; Nuclear Proteins; Pancreatic Neoplasms; Pancreatitis, Chronic; Polymorphism, Single Nucleotide; Prognosis; Retrospective Studies; Risk Factors; Trypsin; Trypsinogen

Pancreatic acinar cells are polarized epithelial cells that store enzymes required for digestion as inactive zymogens, tightly packed at the cell apex. Stimulation of acinar cells causes the zymogen granules to fuse with the apical membrane, and the cells undergo exocytosis to release proteases into the intestinal lumen. Autophagy maintains homeostasis of pancreatic acini. Syntaxin 2 (STX2), an abundant soluble N-ethyl maleimide sensitive factor attachment protein receptor in pancreatic acini, has been reported to mediate apical exocytosis. Using human pancreatic tissues and STX2-knockout (KO) mice, we investigated the functions of STX2 in zymogen granule-mediated exocytosis and autophagy.. We obtained pancreatic tissues from 5 patients undergoing surgery for pancreatic cancer and prepared 80-μm slices; tissues were exposed to supramaximal cholecystokinin octapeptide (CCK-8) or ethanol and a low concentration of CCK-8 and analyzed by immunoblot and immunofluorescence analyses. STX2-KO mice and syntaxin 2. Human pancreatic tissues and dispersed pancreatic acini from control mice exposed to CCK-8 or ethanol plus CCK-8 were depleted of STX2. STX2-KO developed more severe pancreatitis after administration of supramaximal caerulein or a 6-week ethanol diet compared with control. Acini from STX2-KO mice had increased apical exocytosis after exposure to CCK-8, as well as increased basolateral exocytosis, which led to ectopic release of proteases. These increases in apical and basolateral exocytosis required increased formation of fusogenic soluble N-ethyl maleimide sensitive factor attachment protein receptor complexes, mediated by STX3 and STX4. STX2 bound ATG16L1 and prevented it from binding clathrin. Deletion of STX2 from acini increased binding of AT16L1 to clathrin, increasing formation of pre-autophagosomes and inducing autophagy. Induction of autophagy promoted the CCK-8-induced increase in autolysosome formation and the activation of trypsinogen.. In studies of human pancreatic tissues and pancreata from STX2-KO and control mice, we found STX2 to block STX3- and STX4-mediated fusion of zymogen granules with the plasma membrane and exocytosis and prevent binding of ATG16L1 to clathrin, which contributes to induction of autophagy. Exposure of pancreatic tissues to CCK-8 or ethanol depletes acinar cells of STX2, increasing basolateral exocytosis and promoting autophagy induction, leading to activation of trypsinogen.

Topics: Acinar Cells; Animals; Autophagy; Cell Membrane; Ceruletide; Exocytosis; Humans; Mice; Mice, Inbred C57BL; Mice, Knockout; Pancreas; Pancreatic Neoplasms; Pancreatitis; Secretory Vesicles; Syntaxin 1; Trypsinogen

Proteolytic enzymes have shown efficacy in cancer therapy. We present a combination of the two pro-enzymes Trypsinogen and Chymotrypsinogen A with potent in vitro and in vivo anti-tumour efficacy. A synergetic anti-tumour effect for Trypsinogen and Chymotrypsinogen A was determined at a ratio 1:6 (named PRP) using 24 human cancer cell lines. The antiangiogenic effect of PRP was analysed by matrigel-based tube formation and by fibrous capsule formation assays. Furthermore, cell invasion and wound healing assays together with qRT-PCR determination of epithelial-to-mesenchymal transition (EMT) markers were performed on human cancer cells treated with PRP. Additionally, in vivo pharmacokinetic studies were implemented and the PRP's anti-tumour efficacy was explored against orthotopic pancreatic and ovarian cancer tumours. PRP formulation was proven to inhibit in vitro angiogenesis, tumour growth, cancer cell migration and invasiveness; and to be an effective and well tolerated in vivo anti-tumour treatment. Finally, the clinical efficacy of a suppository formulation containing both pancreatic pro-enzymes in the context of a UK Pharmaceuticals Special Scheme was evaluated in advanced cancer patients. Consequently, PRP could have relevant oncological clinical applications for the treatment of advanced or metastatic pancreatic adenocarcinoma and advanced epithelial ovarian cancer.

Topics: Animals; Apoptosis; Cell Proliferation; Chymotrypsinogen; Enzyme Precursors; Female; Humans; Mice; Mice, Nude; Neoplasm Metastasis; Ovarian Neoplasms; Pancreas; Pancreatic Neoplasms; Pilot Projects; Trypsinogen; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Pancreatic juice is a potential source of proteins associated with pancreatic cancer (PC) due to the proximity of ducts to tumor tissue. Therefore, screening of proteins in pancreatic juice from PC patients may identify new PC biomarkers. We analyzed pancreatic juice from patients with pancreatic diseases including PC, chronic pancreatitis (CP) and simple choledocholithiasis (CDS) by 2-DE. Protein spots from PC patients that changed >2-fold compared with both CP and CDS were selected and identified by mass spectrometry (MS). mRNA levels were measured by QRT-PCR in PC cell lines, PC tissues and adjacent pancreatic normal (PN) tissues. Relationships between mRNA levels in PC tissues and their clinical characteristics and promoter methylation were analyzed in PC cell lines and tissues. We found that four proteins were significantly changed in PC compared to CP and simple CDS. Two proteins were up-regulated, serine proteinase-2 (PRSS2) preproprotein and pancreatic lipase-related protein-1 (PLRP1), and two proteins were down-regulated, chymotrypsinogen B (CTRB) precursor and elastase 3B (ELA3B) preproprotein. In all PC cell lines, PRSS 2 mRNA levels were elevated, while PLRP 1 mRNA was detected in 4/5 cell lines. ELA3B mRNA was undetectable in all cell lines, but CTRB mRNA was detected in 2/5 cell lines. In PC tissues compared to PN, levels of PRSS2 mRNA were significantly higher, ELA3B significantly lower, and PLRP1 and CTRB not significantly different. Elevated PRSS2 mRNA levels correlated with high T stage. The ELA3B gene promoter had higher methylation in PC cell lines and tissues compared with PN tissues, and correlated with low ELA3B gene expression. In conclusion, comparative proteomic analysis of pancreatic juice from PC patients is a powerful method to find new PC biomarkers. Hyperexpression of the PRSS2 gene and hypermethylation of ELA3B gene promoter were associated with PC, raising the possibility of their application as new biomarkers in PC diagnosis and screening.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Pancreatic Ductal; Choledocholithiasis; Chymotrypsin; DNA Methylation; Electrophoresis, Gel, Two-Dimensional; Female; Humans; Intracellular Signaling Peptides and Proteins; Male; Middle Aged; Nuclear Proteins; Pancreas; Pancreatic Elastase; Pancreatic Juice; Pancreatic Neoplasms; Pancreatitis, Chronic; Prognosis; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Trypsin; Trypsinogen; Tumor Cells, Cultured

Experimental studies have suggested that trypsinogen may enhance tumor progression and that the ratio between anionic trypsinogen and cationic trypsinogen (HAT/HCT) and between the sum of trypsinogens and pancreatic secretory trypsin inhibitor (PSTI) ((HAT + HCT)/PSTI) are disturbed in patients with pancreatic cancer. The aim of this study was to investigate if pre-diagnostic levels of these parameters are associated with subsequent pancreatic cancer risk.. A total of 33,346 subjects participated in a health screening programme in Malmö, Sweden. Pancreatic cancer cases (n = 84) were matched to three controls each. HAT, HCT and PSTI were analyzed in pre-diagnostic serum samples. Odds ratios for pancreatic cancer were calculated using logistic regression and were then stratified for other risk factors.. In the main analysis, a statistically significant association between the ratio between HAT/HCT and pancreatic cancer was observed for all, for the crude OR and for the ORs adjusted for sex, BMI or Helicobacter pylori. When stratified for sex, statistically significant associations were found for females in the crude OR and for the ORs adjusted for time to analysis, BMI, alcohol consumption or H. pylori. There was a positive association between the ratio of HAT/HCT to pancreatic cancer in the intermediate/high alcohol consumption group and subjects with a BMI <25. The sum of trypsinogens showed a similar pattern, but was only of borderline significance in the intermediate/high alcohol consumption group.. Our hypothesis predicted an increased risk for pancreatic cancer related to an imbalance between trypsin activity and trypsin inhibition capacity. The findings concerning the ratio of HAT/HCT are in line with this. The results related to analyses stratified for other risk factors should be considered as mainly explorative.

Topics: Adult; Alcohol Drinking; Anions; Cations; Female; Humans; Male; Middle Aged; Pancreatic Neoplasms; Prospective Studies; Protein Isoforms; Risk; Smoking; Sweden; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The prevalence and natural history of hereditary pancreatitis (HP) remain poorly documented. The aims of this study were to assess genetic, epidemiological, clinical and morphological characteristics of HP in an extensive national survey.. A cohort comprising all HP patients was constituted by contacting all gastroenterologists and paediatricians (response rate 84%) and genetics laboratories (response rate 100%) in France (60,200,000 inhabitants). Inclusion criteria were the presence of mutation in the cationic trypsingen gene (PRSS1 gene), or chronic pancreatitis in at least two first-degree relatives, or three second-degree relatives, in the absence of precipitating factors for pancreatitis.. 78 families and 200 patients were included (181 alive, 6673 person-years, males 53%, alcoholism 5%, smoking 34%). The prevalence was 0.3/100,000 inhabitants. PRSS1 mutations were detected in 68% (R122H 78%, N29I 12%, others 10%). Penetrance was 93%. Median age at first symptom, diagnosis and date of last news, were 10 (range 1-73), 19 (1-80) and 30 (1-84) years, respectively. HP was responsible for pancreatic pain (83%), acute pancreatitis (69%), pseudocysts (23%), cholestasis (3%), pancreatic calcifications (61%), exocrine pancreatic insufficiency (34%, median age of occurrence 29 years), diabetes mellitus (26%, median age of occurrence 38 years) and pancreatic adenocarcinoma (5%, median age 55 years). No differences in clinical and morphological data according to genetic status were observed. 19 patients died, including 10 directly from HP (8 from pancreatic adenocarcinoma).. The prevalence of HP in France is at least 0.3/100,000. PRSS1 gene mutations are found in 2/3 with a 93% penetrance. Mutation type is not correlated with clinical/morphological expression. Pancreatic adenocarcinoma is the cause of nearly half the deaths.

Topics: Adenocarcinoma; Adolescent; Adult; Age Distribution; Age of Onset; Aged; Aged, 80 and over; Child; Child, Preschool; Epidemiologic Methods; Exocrine Pancreatic Insufficiency; Female; France; Genotype; Humans; Infant; Male; Middle Aged; Mutation; Pancreatic Neoplasms; Pancreatitis, Chronic; Penetrance; Phenotype; Trypsin; Trypsinogen; Young Adult

There is a concept that pancreatitis results from an imbalance of proteases and their inhibitors within the pancreatic parenchyma. It has been recently shown that a loss-of-function variant, c.571G>A (p.G191R), in the anionic trypsinogen (PRSS2) gene protects against chronic pancreatitis in European populations. Here we examined the association of the p.G191R variant with pancreatic disorders in Japan.. Genomic DNA was prepared from 378 healthy controls and 604 patients with pancreatic disorders (241 patients with chronic pancreatitis, 174 with acute pancreatitis, and 189 with pancreatic neoplasm). Mutational analysis of the PRSS2 gene was performed by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing.. The heterozygous p.G191R variant was found in three of 241 (1.2%) patients with chronic pancreatitis, in seven of 174 (4.0%) patients with acute pancreatitis, and in 12 of 189 (6.3%) patients with pancreatic neoplasm. The p.G191R variant was found in 25 (two were homozygous and 23 were heterozygous) of 378 (6.6%) healthy controls. The p.G191R frequency in patients with chronic pancreatitis was lower than that in healthy controls (p = 0.001; odds ratio (OR) 0.178; 95% confidence interval (CI) = 0.057 to 0.561). The p.G191R frequency was lower in patients with alcoholic (0.9%; p = 0.015; OR, 0.132; 95% CI, 0.022 to 0.779) and idiopathic (1.0%; p = 0.025; OR, 0.144; 95% CI, 0.025 to 0.851) chronic pancreatitis than that in healthy controls. There were no statistical differences in the p.G191R frequency between healthy controls and patients with acute pancreatitis or with pancreatic neoplasm. Patients with alcoholic acute pancreatitis (n = 59) had no variant carrier, and the p.G191R frequency was lower than that in healthy controls (p = 0.035).. The p.G191R variant protected against alcoholic and idiopathic chronic pancreatitis as well as alcoholic acute pancreatitis in Japan.

Topics: Adenocarcinoma; Adult; Aged; Alcohol Drinking; Case-Control Studies; DNA Mutational Analysis; Female; Gene Frequency; Genotype; Heterozygote; Homozygote; Humans; Japan; Male; Middle Aged; Multivariate Analysis; Mutation; Odds Ratio; Pancreatic Neoplasms; Pancreatitis; Pancreatitis, Acute Necrotizing; Pancreatitis, Chronic; Trypsin; Trypsinogen

Patients with hereditary pancreatitis (HP) bear a high risk of pancreatic adenocarcinoma, but their life expectancy remains unknown. The objective of the study was to assess whether the high risk of cancer decreases survival.. Inclusion criteria were the presence of a PRSS1 mutation with pancreatic symptoms or chronic pancreatitis in at least two first-degree relatives or three second-degree relatives without another cause. Survival rates were assessed according to risk factors. Excess mortality compared with the general French population was calculated (statistical Esteve model) for two periods (20-50 and 50-70 years), according to several risk factors.. The cohort comprised 189 patients. PRSS1 mutations were found in 66%. A total of 19 patients died at the median age of 60. In all, 10 deaths were attributable to HP, including 8 to pancreatic adenocarcinoma. Median overall survival for the whole cohort was 74 years (95% confidence interval (CI): 71-79). The presence of R122H mutation, gender, tobacco consumption in patients older than 18 years, and diabetes mellitus were not associated with differences in survival. Only patients with pancreatic cancer had decreased survival (P=0.008). Excess mortality risk compared with the general population was 0.02% between 20 and 50 years, and 0.61% between 50 and 70 years (NS). Gender, R122H mutation, diabetes, and tobacco use were not associated with excess mortality in these two periods.. Despite their high risk of cancer, HP patients do not have excess mortality risk compared with the general population, irrespective of gender, tobacco use, or diabetes mellitus. These data should be brought to the patient's attention.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; Female; France; Genetic Predisposition to Disease; Humans; Infant; Male; Middle Aged; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsinogen; Young Adult

An increased risk of pancreatic adenocarcinoma (PA) in patients with hereditary pancreatitis (HP) was previously demonstrated in two multinational studies. The PA frequency in this setting is however unknown due to lack of exhaustive case collection. The aims of this study were to evaluate the standardized incidence ratio (SIR) of PA in an exhaustive national series of patients with HP and to search for risk factors.. All French genetic laboratories (response rate 100%), pediatricians, and gastroenterologists (response rate 84%) were contacted.. mutation in the PRSS1 gene or recurrent, acute, or chronic pancreatitis, with no precipitating factors in two first-degree relatives or >or=3 second-degree relatives in >or=2 generations. Diagnosis of PA was based on histological records.. Seventy-eight families and 200 patients were included (181 alive, 6,673 person-years, median number of generations 3, men 53%, alcoholism 5%, and smoking 34%). PRSS1 mutations were searched for in 96% of the patients and were detected in 68% (maternal inheritance 54%, R122H 78%, N29I 12%, and others 10%). Ten PA were diagnosed (median age 55 yr). SIR of PA for the whole population, men, and women were 87 (95% CI 42-113), 69 (25-150), and 142 (38-225), respectively, with no influence of genetic mutation. At ages 50 and 75 yr, the cumulated risk of PA was 11% and 49% for men and 8% and 55% for women, respectively. Smoking and diabetes mellitus were the main associated risk factors.. Patients with HP have a marked relative and absolute increased risk of PA as compared to the general population, especially in smokers. There is no correlation with the type of PRSS1 mutation.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; DNA; Female; France; Genetic Predisposition to Disease; Humans; Incidence; Infant; Male; Middle Aged; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsinogen

Hereditary pancreatitis should be assumed if other risk factors for the disease can not been identified and if the patient has a family history for recurrent pancreatitis, chronic pancreatitis or pancreatic cancer. Since patients with chronic pancreatitis due to mutations in the cationic trypsinogen-gene have a much higher lifetime risk of developing pancreatic cancer, specifically if they are smokers, an adequate long-term follow up in specialized centers is recommended. The most frequent genetic changes in patients with hereditary pancreatitis are mutations in the cationic trypsinogen gene. Mutations in the CFTR-gene or SPINK1-gene have been reported in patients with idiopathic pancreatitis. The clinical relevance and the therapeutic consequences of these mutations is still controversial. Genetic testing is recommended when a patient with idiopathic pancreatitis is under 25 years at diagnosis or when one or more family members have either pancreatitis or pancreatic cancer. Genetic analysis of asymptomatic family members should only be offered after adequate genetic counselling. Prenatal diagnostic is not recommended.

Topics: Adult; Carrier Proteins; Chromosome Aberrations; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Genes, Dominant; Genetic Counseling; Humans; Pancreatic Neoplasms; Pancreatitis, Chronic; Prognosis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

To investigate mutation of serine protease 1-cationic trypsinogen (CT, PRSS1) gene in members of a Thai family with hereditary pancreatitis and pancreatic cancer.. Polymerase chain reaction and direct sequencing were performed to analyze the PRSS1 gene in two members of the family affected by pancreatitis. Allele specific amplification (ASA) method was then developed to detect the mutation of the PRSS1 gene in all available members of the family and normal control subjects.. A cytosine (C) to thymine (T) mutation at position 2441 (g.2441C>T) of the PRSS1 gene, which results in a substitution of arginine by cysteine at position 116 (R116C) of CT, was identified by direct sequencing in both clinically affected members of the family but was not found in the unaffected member. This mutation, which might be arising from deamination of methylated cytosine in CpG dinucleotide of codon 116 (CGT>TGT), was also detected by the ASA method in the two affected members and a proband's brother but was not observed in unaffected members and 54 normal control subjects.. Autosomal dominant pancreatitis with increased cancer risk in the studied Thai family is most likely due to missense (R116C) mutation in the PRSS1 gene.

Topics: Exons; Family Health; Female; Genetic Predisposition to Disease; Humans; Middle Aged; Mutation, Missense; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Thailand; Trypsin; Trypsinogen

Chronic pancreatitis: Only recently mutations in several genes were found in patients with chronic pancreatitis. In those with a familial chronic pancreatitis mutations of the cationic trypsinogen were identified and the variants N29I and R122H lead to an autosomal dominant disease. In this group of patients the mutation N34S of the trypsin inhibitor SPINK1 was detected. In so-called idiopathic pancreatitis both variants of the SPINK1 and of the CFTR (cystic fibrosis transmembrane conductance regular) were identified. Alterations in both genes were also found in patients with alcoholic chronic pancreatitis. The strongest risk factor for chronic pancreatitis were trypsinogen mutations N29I and R122H mutations. However, both SPINK1 and CFTR increased the risk for chronic pancreatitis to a higher level than alcohol consumption. A genetic investigation should be performed in familial disease and younger age, but also in patients without family history and higher age a mutation could be found. Pancreas cancer: In 10% of the patients with pancreas cancer other members of the family were affected from the disease. Some of them belong to well characterized familial syndroms like HNPCC or Peutz-Jeghers-syndrom. In a minority of the others a genetic factor may be found, too. In sporadic disease the development of the tumor is characterized by continued acquirement of genetic alterations described by the PanIN model (pancreatic intraepithelial neoplesia). This means that the evolution of the neoplasia progresses from normal tissue via epithelial hyperplasy (PanIN 1A), papillary hyperplasy without (PanIN 1B) and with dysplasy (PanIN 2) and carcinoma in situ (PanIN 3) to invasive pancreas cancer. The progression is associated with genetic alterations of the cells (mutations of ki-ras, p16, p53 etc.). This results in deterioration of control of the cell cycle and the apoptosis and explains the malignancy of the disease. These findings may be used in the future to develop newer therapeutic principles in order to improve the dismal prognosis of this disease.

Topics: Carrier Proteins; Cell Transformation, Neoplastic; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Disease Susceptibility; DNA Mutational Analysis; Genes, Dominant; Genotype; Humans; Pancreatic Neoplasms; Pancreatitis; Pancreatitis, Alcoholic; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Mutations in the human cationic trypsinogen are associated with hereditary pancreatitis. The cDNA coding for human cationic trypsinogen was subcloned into the expression vector pcDNA3. The mutations R122H, N29I, A16V, D22G, and K23R were introduced by site directed mutagenesis. We constructed an expression vector coding for active trypsin by subcloning the cDNA of trypsin lacking the coding region for the trypsin activating peptide behind an appropriate signal peptide. Expression of protein was verified by Western blot and measurement of enzymatic activity. AR4-2J cells were transiently transfected with the different expression vectors and cell viability and intracellular caspase-3 activity were quantified. In contrast to wild-type trypsinogen, expression of active trypsin and mutated trypsinogens reduced cell viability of AR4-2J cells. Expression of trypsin and R122H trypsinogen induced caspase-3 activity. Acinar cells might react to intracellular trypsin activity by triggering apoptosis.

Topics: Animals; Apoptosis; Cell Line, Tumor; Cell Survival; Mutagenesis, Site-Directed; Pancreatic Neoplasms; Rats; Recombinant Proteins; Trypsin; Trypsinogen

The protease-activated receptor-2 (PAR-2) is a G protein-coupled receptor that is cleaved and activated by trypsin and tryptase. The purpose of this study was to clarify the role of PAR-2 in proliferation of human pancreatic cancer cells. PAR-2 mRNA and protein expression were detected by RT-PCR and Western blotting in three cell lines, SW1990, Capan-2, and Panc-1. The PAR-2 agonist peptide, SLIGKV (25, 50 micro g/ml) and trypsin (10, 100 ng/ml) significantly increased cell proliferation. Enhancement of MAP kinase also was observed in cancer cells treated with SLIGKV and trypsin. In vivo, subcutaneous xenografted tumors showed significantly enhanced growth after treatment with SLIGKV. Tumor-associated trypsinogen (TAT) mRNA and protein expression was detected in SW1990 and Capan-2, suggesting autocrine trypsin production. PAR-2 activated by trypsin plays an important role in promoting proliferation of pancreatic cancer.

Topics: Animals; Autocrine Communication; Blotting, Western; Cell Division; Humans; Male; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Oligopeptides; Pancreatic Neoplasms; Receptor, PAR-2; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transplantation, Heterologous; Trypsin; Trypsinogen; Tumor Cells, Cultured

The progression of azaserine-induced rat pancreatic adenocarcinoma (AC) was characterised using quantitative and semiquantitative immunohistochemistry for proliferating cell nuclear antigen (PCNA), basement membrane laminin (BML) and trypsinogen (TG). Samples were taken 5-20 months after initiation. High PCNA-labelling indices (PCNA LIs) were measured 5 months after the induction of atypical acinar cell nodules (AACNs), which decreased later and stagnated until a further decline in the month 10 adenomas. Then a second premalignant proliferative wave was observed (month 13) within the adenoma stage. Later, in month 20 differentiated ACs PCNA LIs fell to the host tissue level but were found highest in the month 20 anaplastic ACs indicating a switch to malignant proliferation. Month 20 invasive ACs showed a number of separate proliferative foci. In early AACNs, BML decreased and remained low till the local maximum in the month 13 adenoma. Invasive ACs did not express BML. Month 5 AACN and differentiated AC were TG deficient but anaplastic AC regained its TG expression. However invasive AC was again TG negative. These results are discussed in combination with our previous data on progressional changes of autophagic capacity and microvessel densities.

Topics: Adenocarcinoma; Adenoma; Animals; Azaserine; Biomarkers, Tumor; Carcinogenicity Tests; Cell Count; Disease Progression; Fluorescent Antibody Technique, Indirect; Laminin; Male; Pancreatic Neoplasms; Proliferating Cell Nuclear Antigen; Rats; Rats, Wistar; Trypsinogen

Trypsinogen/trypsin is one of the major serine proteases and is produced by pancreatic acinar cells. Tumor-associated trypsinogen (TAT) has been reported to be produced by several cancer cell lines. The biological roles and activation mechanisms of both TAT and pancreatic acinar trypsinogen (PAT) have not been elucidated in the context of cancer extension, in particular at the stage of invasion and metastasis. In this study, we investigate the roles played by PAT and TAT in pancreatic cancer invasion. In addition, we determined their mechanisms of activation and identified a trypsinogen activity-stimulating factor (TASF) produced by pancreatic cancer cells. TAT expression and high TAT activity were associated with high invasive and liver metastatic potential in SW1990 and CAPAN-2 cells. Moreover, a trypsinogen activating effect and activity prolonging effect was observed in a mixture of these supernatants with trypsinogen. These cells revealed significantly enhanced invasiveness upon invasion assay and in the presence of PAT. TAT and PAT were activated by TASF, active u-PA, produced by pancreatic cancer cells. Activated TAT and PAT can degrade not only ECM proteins but they can also activate other latent proteases. This ECM-protease-network may form a vicious cycle, thereby promoting tumor cell invasion.

Topics: Enzyme Activators; Humans; Immunoblotting; Liver Neoplasms; Neoplasm Invasiveness; Neoplasm Metastasis; Pancreatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; Trypsinogen; Urokinase-Type Plasminogen Activator

Topics: Acute Disease; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Pancreatic Neoplasms; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The bile concentrations of trypsinogen-1, -2 and tumour-associated trypsin-inhibitor (TATI) were determined in 23 patients with benign biliary tract disease, two with biliary tract cancer, and in 15 with pancreatic cancer. We also examined the trypsinogen and TATI expression by immunohistochemistry in tissue specimens from biliary tract cancer and non-neoplastic extrahepatic biliary tract. High levels of trypsinogen-1, trypsinogen-2, and TATI occur in bile of most patients. In contrast to the trypsinogens, the levels of TATI were significantly higher in patients with malignant disease than in those with benign diseases (p=0.04). There was no significant correlation between trypsinogen-2 and amylase (r=0.13, p=0.40), indicating that the occurrence of trypsinogen in bile is not a result of reflux of pancreatic fluid into the bile duct. Immunohistochemically, trypsinogen-2 was detected in five and TATI in 12 out of 15 non-neoplastic biliary tract specimens, and in four and seven out of 11 cholangiocarcinomas, respectively. High concentrations of trypsinogen-1, trypsinogen-2 and TATI occur in the bile of patients with non-neoplastic and malignant biliary tract disease and in patients with pancreatic cancer. At least part of the trypsinogen-2 and TATI found in bile appears to be derived from the biliary epithelium itself.

Topics: Bile; Biliary Tract; Biliary Tract Diseases; Humans; Pancreatic Neoplasms; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Various studies have described increased expression of cationic trypsinogen in malignant tumor cells. To explore the role of secreted cationic trypsinogen in invasion by cancer cells, we introduced cationic trypsinogen cDNA into Panc-1, a pancreatic adenocarcinoma-derived cell line that lacks expression of endogeneous trypsinogen. Four independent clones (designated Panc-1-Try-7, -9, -11 and -24) stably expressing cationic trypsinogen mRNA were isolated and processed for further study. In a zymographic analysis, gelatinolytic activity for cationic trypsinogen was detectable in serum-free conditioned media obtained from all 4 transfectants but not in media from mock-transfected or parental Panc-1 cells. A Matrigel invasion assay revealed that all trypsinogen-expressing transfectants acquired significantly greater invasive ability than that shown by mock-transfected and parental Panc-1 cells. In addition, enhanced invasiveness of the transfectants was suppressed by FUT-175, a serine protease inhibitor, to the level seen in parental cells. These results provide direct evidence that cationic trypsinogen can increase the invasive ability of carcinoma cells.

Topics: Adenocarcinoma; Gelatin; Humans; Immunohistochemistry; Neoplasm Invasiveness; Pancreatic Neoplasms; Transfection; Trypsin; Trypsinogen; Tumor Cells, Cultured

Summary. The understanding of the pathogenesis of chronic pancreatitis is limited. Several theories (i. e. obstruction hypothesis) were suggested in the past but could not be confirmed by experimental data. As a formal description of the course of the disease, the necrosis-fibrosis concept seems to be very attractive. According to this theory, there is no significant difference in the pathogenesis of acute and chronic pancreatitis. A major step was the identification of mutations of the cationic trypsinogen, the secretory trypsin inhibitor (SPINK 1) and the cystic-fibrosis protein (CFTR) in some patients. Investigation of these mutations may significantly contribute to a better understanding of the pathogenesis of chronic pancreatitis.

Topics: Adult; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Fibrosis; Humans; Mutation; Necrosis; Pancreas; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin Inhibitors; Trypsinogen

Hereditary pancreatitis is an autosomal dominant disease. Recently, the genetic defect has been mapped to chromosome 7q35 and consists mainly of a point mutation in exon 3 of the cationic trypsinogen gene which causes an Arg(CGC)-His(CAC) substitution at residue 117. In patients with hereditary pancreatitis the estimated cumulative risk for pancreatic carcinoma to age 70 approaches 40 %. Thus, the role of hereditary pancreatitis in the pathogenesis of pancreatic carcinoma is of interest.. DNA was extracted from peripheral blood (n = 16), fresh tumor tissue (n = 29) and formalin fixed and paraffin embedded tumor tissue (n = 5) of 50 patients with ductal adenocarcinoma of the pancreas. We specifically amplified exon 3 and the intronic flanking sequences of the cationic trypsinogen gene by nested PCR and performed restriction fragment length polymorphism analysis using the restriction enzyme Afl III. In patients with hereditary pancreatitis the G : A point mutation creates a recognition site for Afl III which is not present in unaffected individuals.. None of the 50 patients with ductal adenocarcinoma of the pancreas revealed the G : A point mutation in exon 3 of the cationic trypsinogen gene which is characteristic of hereditary pancreatitis. In addition sequencing of exon 3 did not reveal any other mutations in the DNA of patients with pancreatic adenocarcinoma.. Although hereditary pancreatitis markedly increases the risk for pancreatic cancer, it is rare and probably of little significance with respect to the pathogenesis of the majority of pancreatic adenocarcinomas.

Topics: Adenocarcinoma; Adolescent; Aged; Child; Child, Preschool; DNA; DNA Primers; DNA, Neoplasm; Exons; Humans; Infant; Nucleic Acid Amplification Techniques; Pancreatic Neoplasms; Pancreatitis; Point Mutation; Polymerase Chain Reaction; Risk Factors; Trypsinogen

Vigilin, a protein with a continuous series of 14 KH motifs, forms part of a multiprotein complex containing tRNA. Several lines of evidence have suggested that vigilin expression is enhanced in those cells which were actively engaged in protein synthesis. Accordingly, we show here by immunoelectronmicroscopy a close association of vigilin with the rough endoplasmic reticulum in rat pancreatic cells. Histological examination of these cells furthermore demonstrates the highest intensity of vigilin staining in the perinuclear, intranuclear, and basolateral regions where the endoplasmic reticulum is mainly amassed. In vivo challenge of starving rats fed prior to sacrifice raised in parallel the protein levels of both trypsin and vigilin when compared to unchallenged animals and was associated with enhanced expression of the vigilin gene. In contrast, in human and rat cell lines of pancreatic tumors with a constitutively high expression of vigilin no further stimulation by cholecystokinin treatment could be achieved. Our data provide circumstantial evidence that vigilin may play a crucial role in the ability of an organ, e.g., pancreas, to cope with the physiological demand to upregulate protein synthesis.

Topics: Animals; Carrier Proteins; DNA Primers; Eating; Endoplasmic Reticulum, Rough; Endoplasmic Reticulum, Smooth; Female; Humans; Immunohistochemistry; Microscopy, Immunoelectron; Pancreas; Pancreatic Neoplasms; Polymerase Chain Reaction; Protein Biosynthesis; Proteins; Rats; RNA-Binding Proteins; Trypsin; Trypsinogen; Tumor Cells, Cultured; Uterus

A 71-year-old woman was admitted with the suspected diagnosis of pancreatic carcinoma. As a child she had had repeated attacks of abdominal pain of undetermined cause. When aged 48 years she had developed diabetes mellitus. Her now 42-year-old daughter had from the age of 9 years suffered from repeated attacks of acute pancreatitis that had finally led to chronic pancreatitis. The patient's 15-year-old grandchild was having recurrent bouts of abdominal pain.. Imaging procedures revealed calcifications in the pancreas and an infiltrating space-occupying lesion, about 3 cm in diameter, in the head of the pancreas with lymph node and liver metastases. Cytological analysis of material aspirated from the space-occupying mass showed typical findings of ductal pancreatic carcinoma. FURTHER TESTS, TREATMENT AND COURSE: At first the patient's course was not typical for a genetically-determined disease, but the family history raised the suspicion of hereditary pancreatitis. A genetic test (Afl-III-RFLP test) demonstrated the mutation Arg 117 His in the cationic trypsinogen gene in all diseased or symptomatic family members. The patient died of the complications of the pancreatic cancer.. Genetic tests are valuable in the diagnosis of hereditary pancreatitis, because the increased cancer risk can be met by frequent examinations in affected family members.

Topics: Abdominal Pain; Acute Disease; Adolescent; Adult; Aged; Calcinosis; Chronic Disease; Family; Fatal Outcome; Female; Humans; Pancreas; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Point Mutation; Polymorphism, Restriction Fragment Length; Recurrence; Risk Factors; Tomography, X-Ray Computed; Trypsinogen; Ultrasonography

Topics: Carcinoma, Acinar Cell; Carcinoma, Neuroendocrine; Combined Modality Therapy; Fatal Outcome; Humans; Immunoenzyme Techniques; Liver Neoplasms; Male; Microscopy, Immunoelectron; Middle Aged; Neoplasms, Multiple Primary; Pancreatic Neoplasms; Skin Neoplasms; Trypsinogen

We examined whether human pancreatic ductal cancer cells express and secrete pancreatic cationic trypsinogen in vitro which can be spontaneously converted into active trypsin at acidic pH (pH 4.5-5. 5), in contrast to anionic trypsinogen. Cationic trypsinogen expression at the mRNA level was observed in differentiated Capan-1 and BxPC-3 cell lines. However, expression was not detected in either poorly-differentiated Panc-1 or undifferentiated MIAPaCa-2 cell line. The gelatinolytic activity of the activated form of trypsinogen in each conditioned medium in the presence of enterokinase (1.0 microg/ml) (a band with a molecular weight of approximately 23 kDa) corresponded well to the level of cationic trypsinogen mRNA. The spontaneous activation of trypsinogen also was observed by gelatin zymography of the acid-loaded conditioned medium (pH 5.5). These findings suggest that trypsinogen produced by human pancreatic ductal cancer has the characteristics of spontaneous activation and gelatinolytic activity in the presence of proton.

Topics: Culture Media, Conditioned; Gelatin; Gels; Humans; Hydrogen-Ion Concentration; Pancreatic Ducts; Pancreatic Neoplasms; Trypsin; Trypsinogen; Tumor Cells, Cultured

Serum concentrations of trypsinogen-2 and trypsin-2-alpha(1)-antitrypsin (trypsin-2-AAT) were determined in 145 patients with malignant and 61 with benign digestive-tract diseases. The validity of these tests for detection of cancer was compared with that of CA 19-9 and CEA. Elevated levels of trypsinogen-2 (>90 micrograms/l) and trypsin-2-AAT (>25 micrograms/l) were found in 46% and 42%, respectively, of patients with malignant disease and the levels of trypsinogen-2 were significantly higher than in those with benign disease (p<0.005). High trypsinogen-2 and trypsin-2-AAT concentrations were found most often in patients with biliary and pancreatic cancer, but also in benign obstructive biliary disease. Our results suggest that trypsinogen-2 and trypsin-2-AAT are new potential markers for cholangiocarcinomas.

Topics: alpha 1-Antitrypsin; Amylases; Bile Duct Neoplasms; Bilirubin; Biomarkers; Biomarkers, Tumor; Carcinoma, Hepatocellular; Cholangiocarcinoma; Digestive System Diseases; Digestive System Neoplasms; False Positive Reactions; Humans; Liver Neoplasms; Pancreatic Neoplasms; Trypsin; Trypsinogen

Topics: Alcohol Drinking; Biomarkers; Enzyme-Linked Immunosorbent Assay; Humans; Pancreatic Diseases; Pancreatic Neoplasms; Trypsin; Trypsinogen

To test the diagnostic utility of pancreatic digestive enzyme immunohistochemistry in liver cancers, the expression of three pancreatic digestive enzymes (trypsinogen, chymotrypsinogen and pancreatic lipase) was investigated in cholangiocarcinoma (CC) (n = 42), hepatocellular carcinoma (HCC) (n = 35), combined HCC-CC (n = 11) and metastatic adenocarcinoma (MA) of the liver (n = 34; 4 gastric cancer, 5 pancreatic cancer and 25 colon cancer). In CC, 15 (36%) expressed one or more of these enzymes, while the remaining 27 (64%) did not express any enzymes. In MA, 13 (38%) expressed one or more of these enzymes, while the remaining 21 (62%) did not express any enzymes. Expression of trypsinogen, chymotrypsinogen and lipase was noted in 15 CC (36%), 11 CC (25%) and 15 CC (36%), respectively, and in 9 MA (26%), 6 MA (18%) and 13 MA (38%), respectively. There was no significant difference in the positive ratio of each enzyme between CC and MA. In positive cases, the enzymes were expressed with a cytoplasmic granular pattern. In MA, there was no significant difference in the positive ratio of the enzymes among the primary sites. In contrast to CC and MA, these enzymes were not expressed in any cases of HCC and combined HCC-CC. These data suggest that pancreatic digestive enzyme immunohistochemistry may be useful for differential diagnosis between HCC and CC or MA as well as between combined HCC-CC and CC or MA, but it is not useful for differential diagnosis between CC and MA. A positive reaction for these enzymes is indicative of CC or MA and is against the diagnosis of HCC or combined HCC-CC, and a negative reaction is noncontributory to the differential diagnosis.

Topics: Adenocarcinoma; Aged; Carcinoma, Hepatocellular; Cholangiocarcinoma; Chymotrypsinogen; Colonic Neoplasms; Diagnosis, Differential; Female; Humans; Immunoenzyme Techniques; Liver Neoplasms; Male; Middle Aged; Pancreas; Pancreatic Neoplasms; Pancrelipase; Stomach Neoplasms; Trypsinogen

We previously demonstrated that two human pancreatic adenocarcinoma cell lines, CFPAC-1 (established from a patient with cystic fibrosis) and CAPAN-1, were able to secrete trypsinogens 1 and 2 specifically. In order to analyze the relation of trypsin secretion to differentiation and cell growth, we undertook a comparative study of immunoreactive trypsin 1 (IRT) secretion by the two cell lines during cell growth in the presence and in the absence of various differentiating agents: sodium butyrate (NaBut), dimethylsulfoxide (DMSO), and dexamethasone (DX). In the presence of NaBut, IRT levels in the supernatants of both cell lines were slightly increased, whereas the cellular growth of both cell lines decreased significantly. In the presence of DX, IRT levels in cell culture conditioned media immediately and dramatically decreased, but the cell growth of neither cell line was affected by DX. An important increase in IRT levels was observed when CFPAC-1 cells and CAPAN-1 cells were grown in the presence of DMSO, but for both cell lines the cellular growth decreased in the presence of DMSO. Our data show that neither the IRT secretion level nor the differentiation state of these cell lines correlates with cellular growth, and suggests that the expression of pancreatic proteases by these two tumor cell lines could be either related to a common stem cell with this potential or to a possible acinar origin of pancreatic cancer, as recently proposed by others.

Topics: Adenocarcinoma; Butyrates; Butyric Acid; Cell Division; Dexamethasone; Dimethyl Sulfoxide; Humans; Pancreatic Neoplasms; Trypsin; Trypsinogen; Tumor Cells, Cultured

Expression of pancreatic trypsinogen and cathepsin B in 23 surgically resected pancreatic ductal adenocarcinomas was evaluated immunohistochemically, using a monoclonal antibody against human pancreatic trypsinogen and a polyclonal antibody against human cathepsin B. Fifteen of 20 invasive tubular adenocarcinomas (75%) expressed pancreatic trypsinogen in a coarse granular pattern located in the supranuclear cytoplasm of the carcinoma cells. In addition, metastatic lesions, including those in peripancreatic lymph nodes and neural plexuses, expressed pancreatic trypsinogen. In contrast, three intraductal (non-invasive) papillary adenocarcinomas did not express pancreatic trypsinogen. Cathepsin B expression was recognised in 14 of 20 invasive tubular adenocarcinomas (70%) in a fine granular pattern located diffusely in the cytoplasm of the carcinoma cells, while none of the three intraductal papillary adenocarcinomas had detectable cathepsin B. These findings suggest that pancreatic invasive ductal adenocarcinomas express pancreatic trypsinogen and cathepsin B immunoreactive peptides, raising the possibility that pancreatic trypsinogen and cathepsin B may act independently of each other in the process of carcinoma invasion and metastasis, like other different classes of proteases involved in the proteolytic modification of the matrix barrier.

Topics: Adult; Aged; Antibodies, Monoclonal; Antibody Specificity; Carcinoma, Ductal, Breast; Cathepsin B; Female; Humans; Immunohistochemistry; Lymphatic Metastasis; Male; Middle Aged; Pancreatic Neoplasms; Trypsinogen

Proteins with trypsin-like immunoreactivity (first detected by a specific immunoenzymatic assay) were isolated from CAPAN-1 and CFPAC-1 cell culture-conditioned media by chromatography on an immunoadsorbent prepared with a polyclonal antibody directed against trypsin 1. The adsorbed proteins were devoid of free trypsin activity but trypsin activity was present after enterokinase activation demonstrating that the immunoreactive trypsin present in cell supernatants corresponds to trypsinogens. When characterised by Western blotting using a monoclonal antibody directed against human trypsin 1 two protein bands corresponding to trypsinogen 1 (23 kDa) and trypsinogen 2 (25 kDa) gave a positive reaction. These results demonstrate the presence of trypsinogens 1 and 2 in CAPAN-1 and CFPAC-1 cells and in their culture-conditioned media.

Topics: Adenocarcinoma; Blotting, Western; Cell Line; Enteropeptidase; Enzyme Activation; Humans; Molecular Weight; Pancreatic Neoplasms; Trypsin; Trypsinogen; Tumor Cells, Cultured

The serum behavior of amylase, pancreatic isoamylase, lipase, trypsinogen, and elastase 1 was studied in 145 patients with pancreatic disease and in 66 patients with abdominal pain of nonpancreatic origin, for the purpose of evaluating the relative diagnostic utility of their assays. In 34 patients with acute pancreatitis, serum lipase, trypsinogen, and elastase 1 were elevated in all 34, pancreatic isoamylase in 33 (97%) and amylase in 30 (88%). Ten of these acute pancreatitis patients were followed sequentially for seven days: the variations in their serum enzyme levels were parallel, although the lipase, trypsinogen, and particularly the elastase 1 elevations persisted longer than did those of amylase and pancreatic isoamylase. Among the patients with chronic pancreatitis, either in painful relapse (N = 19) or with pancreatic cysts (N = 15), the respective percentages of enzymes elevations were: 79 and 80% for elastase 1, 68 and 67% for trypsinogen, 63 and 73% for pancreatic isoamylase, 58 and 60% for lipase, 53 and 60% for amylase. In the 52 chronic pancreatitis patients studied during clinical remission, serum enzyme behavior varied greatly, and a majority of the assays (60%) were normal; even in the case of severe pancreatic exocrine insufficiency, normal as well as abnormally high and low enzyme values were seen. Highly variable enzyme behavior was also seen in the 40 patients with pancreatic cancer, and elastase I was the most frequently (35%) elevated enzyme in this group as well. Among the patients with abdominal pain of nonpancreatic origin, abnormally high enzyme levels were present in percentages ranging from 6% for lipase to 21% for trypsinogen.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acute Disease; Adult; Aged; Amylases; Chronic Disease; Clinical Enzyme Tests; Female; Humans; Isoamylase; Lipase; Male; Middle Aged; Pancreatic Diseases; Pancreatic Elastase; Pancreatic Neoplasms; Trypsinogen

A simple method for the purification of anionic and cationic trypsinogen and trypsin from human pancreatic juice applying affinity chromatography on aprotinin coupled Sepharose is described together with the N-terminal amino acid sequences for both trypsinogens. In addition, enzyme-linked immunoabsorbent assay (ELISA) methods for the determination of anionic and cationic trypsin-like immunoreactivity (irAT and irCT) are described. Normal serum levels are 21.3 +/- 7.4 micrograms/l and 27.8 +/- 9.0 microgram/l for irAT and irCT respectively and the accuracy of these assays is 6-10%. In our population, the normal ratio between irCT and irAT in serum is 1.36 +/- 0.42. In normal serum trypsin-like immunoreactivity consists solely of trypsinogen. In acute pancreatitis there is an increase over normal of both irAT and irCT with a proportionally greater increase in irAT than irCT. Similar changes are also found in uremic patients.

Topics: Acute Disease; Amino Acid Sequence; Anions; Cations; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Humans; Kidney Diseases; Molecular Sequence Data; Molecular Weight; Pancreatic Juice; Pancreatic Neoplasms; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

Immunoreactive trypsin in serum can be divided into trypsinogen and trypsin-alpha 1-proteinase inhibitor (alpha 1PI) complexes. These were studied separately in serum from 204 patients with acute gastro-intestinal symptoms. Elevated levels of both trypsinogen and trypsin-alpha 1PI complexes were seen in patients with acute pancreatitis. Elevated levels of trypsinogen and normal or slightly elevated levels of trypsin-alpha 1PI complexes were seen in patients with biliary tract diseases. An isolated increase in the concentration of trypsin-alpha 1PI complexes with normal trypsinogen and amylase levels were seen in patients with perforated ulcer. This third cluster may result from an absorption of active trypsin from the peritoneal cavity. Small amounts of trypsin-alpha 1PI complexes were present also in serum from patients free from pancreatic disease. The results in this study show that high levels of trypsin-alpha 1PI complexes in serum are seen mainly in patients with acute pancreatitis. However, elevated levels are also seen in other pathological conditions in the upper gastrointestinal tract; therefore an assay for these complexes is not a specific diagnostic test for acute pancreatitis.

Topics: Abdomen, Acute; alpha 1-Antitrypsin; Biliary Tract Diseases; Gastritis; Humans; Pancreatic Neoplasms; Pancreatitis; Peptic Ulcer; Trypsinogen

Treatment of AR4-2J cells with dexamethasone at 10 nM for 96 h inhibited cell replication by 75% and increased cell size (30%), protein content (1.6-fold) and protein synthesis (2-fold). The increase in protein synthesis was largely due to a 5 to 10-fold increase in the synthesis of secretory proteins. Amylase activity increased 20 to 30-fold in cellular homogenates and 10 to 20-fold in culture medium. Both in the presence and absence of dexamethasone AR4-2J cells release their secretory proteins by constitutive secretion. The proportion of newly synthesized amylase retained by the cells over the 14 h labeling period increased from 15 to 30% with hormone treatment. As judged by comigration on polyacrylamide gels and Western blots analyzed by immunospecific sera, AR4-2J cells synthesize and secrete the majority of known pancreatic secretory proteins. Dexamethasone increased the synthesis of trypsinogen 12 to 16-fold, chymotrypsinogen 4.5 to 6-fold, the group of procarboxypeptidases 6-fold, and amylase 7 to 10-fold. Messenger RNA levels for trypsinogen, amylase and lipase were each increased 4 to 5-fold. At the ultrastructural level dexamethasone led to significant increases in rough endoplasmic reticulum (RER) (30-fold) and Golgi elements (1.5-fold) and to the de novo appearance of electron-opaque granules (0.1-0.5 microns) which were shown to contain amylase by immunolocalization techniques employing protein A-gold. Dexamethasone also led to the formation of gap junctions between AR4-2J cells. These findings indicate that AR4-2J cells provide a model for differentiation of pancreatic acinar cells which should also be studied for the differentiation markers for the regulated secretory pathway.

Topics: Amylases; Animals; Carboxypeptidases; Carcinoma; Cell Line; Chymotrypsinogen; Cytoplasmic Granules; Endoplasmic Reticulum; Glucocorticoids; Golgi Apparatus; Microscopy, Electron; Neoplasm Proteins; Pancreatic Neoplasms; Rats; Trypsinogen; Tumor Cells, Cultured

In order to investigate the role of renal factors in affecting trypsinogen 1 metabolism and excretion in chronic pancreatic disease, serum immunoreactive trypsin (IRT), urinary IRT, gamma-glutamyltransferase (GGT), alpha-glucosidase (AGL) and RNase outputs and the molecular size distribution of serum and urine IRT were studied in 8 control subjects, 18 cases with pancreatic cancer, and 23 cases with chronic pancreatitis. Serum chromatography demonstrated that most immunoreactivity eluted as trypsinogen 1. Smaller amounts of immunoreactivity at higher molecular weights were also observed. Urine chromatography displayed both trypsinogen 1 and heavier molecular forms. An inverse linear correlation was noticed between creatinine clearance and serum trypsinogen 1 levels. Multiple regression analysis (urinary IRT output dependent and GGT, AGL, and RNase predictor variables) showed a significant linear correlation. RNase was found to be the most important parameter in explaining urinary IRT output. Mild variations in the glomerular function seem to be able to influence serum trypsinogen 1 levels. Urinary IRT excretion is principally explained by a disturbance in the tubular reabsorption of low molecular weight proteins, such as RNase.

Topics: Adult; Aged; Chronic Disease; Female; Humans; Kidney; Male; Middle Aged; Pancreatic Diseases; Pancreatic Neoplasms; Trypsin; Trypsinogen

In order to investigate the role of circulating free trypsinogen and renal tubular dysfunction in affecting trypsin plasma-urine transfer, serum immunoreactive trypsin (IRT), its urinary output, IRT molecular size distribution, filtrable immunoreactive trypsin, gamma-glutamyltransferase and alpha-glucosidase outputs were studied in 6 control subjects, 9 patients with pancreatic cancer and 15 with chronic pancreatitis. The majority of immunoreactivity was always eluted at a molecular weight of about 24,000 and might therefore be considered as free trypsinogen. Variable amounts of IRT at higher molecular weights, possibly represented by trypsin-inhibitor complexes, were also detected. Increasing IRT levels were generally accounted for by free trypsinogen, regardless of the nature of the disease. Unlike serum free trypsinogen levels, renal tubular damage, evaluated by means of the excretion of two high-molecular weight urinary enzymes, seems to play a prominent role in explaining trypsin plasma-urine transfer.

Topics: Adolescent; Adult; Aged; alpha-Glucosidases; Carcinoma, Intraductal, Noninfiltrating; Chronic Disease; Female; gamma-Glutamyltransferase; Humans; Kidney Diseases; Kidney Tubules; Male; Metabolic Clearance Rate; Middle Aged; Molecular Weight; Pancreatic Neoplasms; Pancreatitis; Trypsin; Trypsinogen

Small, but significant amounts of immunoreactive trypsin/trypsinogen (IRT) are excreted in bile, obtained via percutaneous transhepatic catheter in patients with complete distal bile duct obstruction, and thus uncontaminated with pancreatic juice. The major serum proteolytic enzyme inhibitors alpha 2-macroglobulin and alpha 1-antitrypsin are also present in very small amounts in bile. The bile-to-serum ratios of these inhibitors are much lower (approximately two orders of magnitude) than that for IRT. The role of these inhibitors and their relative importance in biliary proteolytic enzyme inhibition is unknown.

Topics: Adenoma, Bile Duct; Aged; Bile; Common Bile Duct Neoplasms; Humans; Male; Middle Aged; Pancreatic Neoplasms; Radioimmunoassay; Trypsin; Trypsinogen

Previous studies, in selected populations, have determined that a low serum trypsinogen can be seen in chronic exocrine pancreatic disorders (CP) and primary diabetes mellitus (DM). In this study, we investigated the predictive value of a low serum trypsinogen. The study population consisted of 488 consecutive emergency room patients admitted to our hospital on whom a serum amylase was drawn by the emergency room staff. Of the sera drawn, 418 were saved and tested for immunoassayable trypsinogen. Ten of 418 (2.4%), had a low level of this marker (less than 10 ng/ml). Of these 10, four had obvious historical or clinical evidence of CP during their initial hospitalization. Six patients, however, had no initial evidence of CP. Follow-up was obtained in three of the six, and all three had evidence of CP despite absence of symptoms. Of the 418 patients, 37 had DM. A low trypsinogen was found in three of these 37, and all three had concomitant CP. We conclude that this new assay has excellent predictive value in diagnosing chronic exocrine pancreatic disorders.

Topics: Adult; Aged; Chronic Disease; Diabetes Mellitus; Emergency Service, Hospital; Female; Humans; Male; Middle Aged; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Immunoreactive trypsinogen and pancreatic secretory trypsin inhibitor (PSTI) were demonstrated in pancreas by means of an immunoperoxidase technique. They had the same distribution in acinar cells of 'normal' human exocrine pancreas tissues. Ductal adenocarcinoma tissue and pancreatic undifferentiated carcinoma contained neither antigen. Scattered 'normal'-looking cells in the border area between normal and neoplastic tissue of both types of tumor stained positively for trypsinogen and for PSTI.

Topics: Adenocarcinoma; Carcinoma; Humans; Immunoenzyme Techniques; Pancreas; Pancreatic Neoplasms; Trypsin Inhibitor, Kazal Pancreatic; Trypsin Inhibitors; Trypsinogen

We have previously established that secretory proteins from a rat acinar cell tumor lack two forms of procarboxypeptidase B, are deficient in a major lipase species, and possess markedly reduced amounts of the basic proteins proelastase, basic chymotrypsinogen, basic trypsinogen and ribonuclease (Iwanij, V., and J.D. Jamieson, J. Cell Biol., 95:734-741). Because secretory proteins are markers for acinar cell differentiation, we sought to establish whether the secretory protein profile of the acinar cell tumor is unique to the transformed cell or whether it resembles that of a stage of normal pancreatic development. To this end, we compared the secretory protein pattern from acinar tumor cells with that of rat pancreatic rudiments at days 19-22 of gestation and through day 21 of the postnatal period. Two-dimensional IEF-SDS gel electrophoresis coupled with biosynthetic labeling and fluorography indicates a time-dependent appearance of individual secretory proteins with basic polypeptides, except for amylase, appearing in the terminal stages of differentiation. In comparison, the secretory protein pattern of the acinar tumor cells most closely resembles that of day-19 embryonic pancreatic rudiments. We propose that the cells of the acinar cell tumor may, in part, mirror a stage of normal pancreatic development.

Topics: Amylases; Animals; Carboxypeptidase B; Carboxypeptidases; Cell Differentiation; Chymotrypsinogen; Enzyme Precursors; Hydrolases; Lipase; Pancreas; Pancreatic Elastase; Pancreatic Neoplasms; Rats; Rats, Inbred Strains; Ribonucleases; Trypsinogen

Topics: alpha-Amylases; Endoribonucleases; Follow-Up Studies; Humans; Kidney; Kinetics; Pancreas; Pancreatectomy; Pancreatic Hormones; Pancreatic Juice; Pancreatic Neoplasms; Saliva; Trypsinogen

Topics: Adult; Aged; Female; Gallstones; Hepatitis, Alcoholic; Humans; Male; Middle Aged; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

Serum immunoreactive trypsinogen (IT) levels were measured in 479 normal controls and in 604 patients (510 with nonpancreatic diseases and 94 with pancreatic diseases) in order to evaluate the distribution of IT values in the control population and the accuracy of the assay in the diagnosis of pancreatic diseases. It concentrations were normally distributed in the healthy population; children showed mean IT values significantly lower than adults. The sensitivity, specificity, and predictive value of a positive and negative result in diagnosing acute or chronic pancreatitis were evaluated vs normals and vs normals plus all patients. With an IT value higher than 80 ng/ml, the likelihood that a patient is not affected by acute pancreatitis is less than 5%. An IT value lower than 9 ng/ml detected 44% of chronic pancreatitis and was related to a 52% probability of such a condition. The 48% false positive results also include patients with pancreatic tumor (31% of cases), so that the chance of finding reduced IT levels in subjects without pancreatic damage drops to 17%. In view of the low prevalence of pancreatic diseases, IT assay should not be taken into consideration as a diagnostic screening test in the general population and its use should be limited to a hospitalized population.

Topics: Adolescent; Adult; Aged; Cations; Child; Child, Preschool; False Positive Reactions; Female; Humans; Infant; Male; Middle Aged; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Radioimmunoassay; Trypsinogen

The transplantable pancreatic acinar carcinomas of rat established recently provide useful model systems to examine the composition of secretory proteins as well as the secretory process in transformed pancreatic exocrine epithelium. The neoplastic acinar cells exhibit considerable variation in the extent of cytodifferentiation. In the present study the enzymatic profile of this heterogeneous tumor cell population has been investigated by the indirect immunofluorescent technique using antibodies against six pancreatic enzymes. By immunofluorescence, all neoplastic cells stained positively for the six enzymes tested: amylase, lipase, carboxypeptidase A, chymotrypsinogen, trypsinogen, and ribonuclease. Some variability in the intensity of immunofluorescence was noted, suggesting possible quantitative differences in the content of a given enzyme among tumor cells. These observations suggest that neoplastic acinar cells with or without secretory granules contain secretory proteins, but to a variable extent.

Topics: Amylases; Animals; Carboxypeptidases; Carboxypeptidases A; Chymotrypsinogen; Epithelium; Fluorescent Antibody Technique; Immunodiffusion; Lipase; Pancreas; Pancreatic Neoplasms; Rats; Ribonucleases; Trypsinogen

Polyacrylamide gel electrophoresis of pure pancreatic juice from 14 healthy normal subjects, 11 chronic alcoholics without detectable pancreatic disease, 15 patients with pancreatitis, and two with cancer of the pancreas consistently demonstrated the presence of two variants of trypsinogen with different electrophoretic mobilities. In healthy normal subjects the proportion of cationic to anionic trypsinogen was invariably greater than 1 and averaged about 2. In chronic alcoholics, patients with pancreatitis or cancer of the pancreas, this ratio, with a single exception, was below one and averaged about 0.45. The extraordinary consistency of these findings suggests that the quantitative relationship between cationic and anionic trypsinogen in human pancreatic juice may be a very sensitive indicator of incipient or existing pancreatic pathology. The most acceptable explanation for the reversal of the normal zymogen ratio in pancreatic disease is a selective increase in the synthesis of the anionic variant relative to that of the cationic species. Total trypsinogen concentrations differed widely from one another in the three patient groups, but the ratio of cationic to anionic trypsinogen exhibited little change and remained below 1. Our results also demonstrate for the first time a specific effect of chronic alcohol abuse on the secretory profile of a pancreatic enzyme in human subjects. A newly discovered minor, trypsinogen-like component of human pancreatic juice was found to be significantly increased in pancreatic juice of chronic alcoholics, decreased in pancreatic secretions of patients with pancreatitis, and barely detectable in those of two patients with cancer of the pancreas.

Topics: Adult; Alcoholism; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Pancreatic Juice; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Pancreatic carcinomas electively induced by immunological mechanism [(1), (2)] keep their exocrine secretory specificity along the various stages of their evolution: (a) during the transformation phase from adenoma to carcinoma; (b) in the evolved carcinoma; (c) in its metastasis; (d) in the ascitic carcinomatous cells formed. They are called: immuno-inducted carcinoma. The carcinomatous cells of the constantly deadly ascites cease their production of secretion granules after passages by intraperitoneal graft; but this secretion reappears in the solid carcinomas they induce by subcutaneous graft and contains trypsinogen and chymotrypsinogen, even after the 92nd passage, at the 757 day. Besides, the antisera (1) enhance the growth and the affinity for pancreas and adipose tissue of the carcinomatous ascitic strains they induced. They, sometimes, produce nodular hepatic carcinomas.

Topics: Antibodies; Ascites; Carcinoma; Chymotrypsinogen; Lipoprotein Lipase; Lymph Nodes; Lymphatic Metastasis; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Pancreatic Neoplasms; Trypsinogen

Topics: Amylases; Child; Cystic Fibrosis; Enteropeptidase; Humans; Infant; Methods; Neutropenia; Pancreas; Pancreatic Cyst; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Peptide Hydrolases; Syndrome; Trypsinogen

Topics: Bile; Enzyme Activation; Humans; In Vitro Techniques; Pancreatic Neoplasms; Trypsinogen

Topics: Animals; Carboxypeptidases; Cattle; Chymotrypsin; Chymotrypsinogen; Enzyme Activation; Enzyme Precursors; Humans; Male; Middle Aged; Pancreatic Ducts; Pancreatic Elastase; Pancreatic Juice; Pancreatic Neoplasms; Peptide Hydrolases; Phospholipases; Proteins; Time Factors; Trypsin; Trypsin Inhibitors; Trypsinogen