A full genome scan for gastric cancer.
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Environment↗
Biomedical subjects
T Minamoto
Publications and source records attributed to T Minamoto.
Distinct pattern of p53 phosphorylation in human tumors.
The protein product of the tumor suppressor gene p53 is phosphorylated on multiple residues by several protein kinases. Using a battery of 10 antibodies developed against different phosphorylated and acetylated residues of p53, we compared the pattern of p53 phosphorylation and acetylation in tumor-derived cell lines, tumor samples, and non-neoplastic cells. Irrespective of tumor types or the presence of p53 mutation, phosphorylation and acetylation of p53 was substantially higher in samples obtained from tumor tissues than those found in non-transformed samples. Among the 10 sites analysed, phosphorylation of residues 15, 81, 392, and acetylation were among the more frequent modifications. Analysis of two of the more abundant phosphorylation or acetylation sites on p53 is sufficient to detect 72% of tumor-derived p53 proteins. The distinct pattern of p53 phosphorylation and acetylation in human tumors may offer a new means to monitor the status and activity of p53 in the course of tumor development and progression.
Acetylation↗
Gene mutation as a target for early detection in cancer diagnosis.
The increasing number of genetic aberrations implicated in the development of human cancer has prompted a search to detect them at the earliest possible stage of their formation. Of the many such genetic changes identified thus far, relatively few meet the standard for markers in early diagnosis and prognosis, namely that the genetic modifications occur during the early onset phase of cancer development. Parallel to the increasing number of such genes is the growing availability of technologies using more powerful and cost-efficient methods that enable mass screening for genetic alterations. The purpose of this review is to summarize the currently available genes that can serve as markers for early detection of cancers and methods that allow their detection.
DNA Methylation↗
Endosonographic Features of advanced gastric carcinoma invading the colon: is it easy to differentiate from pancreatic invasion?
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Colon↗
Jun NH2-terminal kinase phosphorylation of p53 on Thr-81 is important for p53 stabilization and transcriptional activities in response to stress.
The p53 tumor suppressor protein plays a key role in the regulation of stress-mediated growth arrest and apoptosis. Stress-induced phosphorylation of p53 tightly regulates its stability and transcriptional activities. Mass spectrometry analysis of p53 phosphorylated in 293T cells by active Jun NH2-terminal kinase (JNK) identified T81 as the JNK phosphorylation site. JNK phosphorylated p53 at T81 in response to DNA damage and stress-inducing agents, as determined by phospho-specific antibodies to T81. Unlike wild-type p53, in response to JNK stimuli p53 mutated on T81 (T81A) did not exhibit increased expression or concomitant activation of transcriptional activity, growth inhibition, and apoptosis. Forced expression of MKP5, a JNK phosphatase, in JNK kinase-expressing cells decreased T81 phosphorylation while reducing p53 transcriptional activity and p53-mediated apoptosis. Similarly transfection of antisense JNK 1 and -2 decreased T81 phosphorylation in response to UV irradiation. More than 180 human tumors have been reported to contain p53 with mutations within the region that encompasses T81 and the JNK binding site (amino acids 81 to 116). Our studies identify an additional mechanism for the regulation of p53 stability and functional activities in response to stress.
Animals↗
Analysis of JNK, Mdm2 and p14(ARF) contribution to the regulation of mutant p53 stability.
Identification of Mdm2 and JNK as proteins that target degradation of wt p53 prompted us to examine their effect on mutant p53, which exhibits a prolonged half-life. Of five mutant p53 forms studied for association with the targeting molecules, two no longer bound to Mdm2 and JNK. Three mutant forms, which exhibit high expression levels, showed lower affinity for association with Mdm2 and JNK in concordance with greater affinity to p14(ARF), which is among the stabilizing p53 molecules. Monitoring mutant p53 stability in vitro confirmed that, while certain forms of mutant p53 are no longer affected by either JNK or Mdm2, others are targeted for degradation by JNK/Mdm2, albeit at lower efficiency when compared with wt p53. Expression of wt p53 in tumor cells revealed a short half-life, suggesting that the targeting molecules are functional. Forced expression of mutant p53 in p53 null cells confirmed pattern of association with JNK/Mdm2 and prolonged half-life, as found in the tumor cells. Over-expression of Mdm2 in either tumor (which do express endogenous functional Mdm2) or in p53 null cells decreased the stability of mutant p53 suggesting that, despite its expression, Mdm2/JNK are insufficient (amount/affinity) for targeting mutant p53 degradation. Based on both in vitro and in vivo analyses, we conclude that the prolonged half-life of mutant p53 depends on the nature of the mutation, which either alters association with targeting molecules, ratio between p53 and targeting/stabilizing molecules or targeting efficiency.
Cell Membrane↗
Wnt/beta-catenin signaling induces the expression and activity of betaTrCP ubiquitin ligase receptor.
Beta-transducing repeat-containing protein (betaTrCP) targets the ubiquitination and subsequent degradation of both beta-catenin and IkappaB, thereby playing an important role in beta-catenin/Tcf and NF-kappaB-dependent signaling. Here evidence is presented that beta-catenin/Tcf signaling elevates the expression of betaTrCP mRNA and protein in a Tcf-dependent manner, which does not require betaTrCP transcription. Induction of betaTrCP expression by the beta-catenin/Tcf pathway results in an accelerated degradation of the wild-type beta-catenin, suggesting that the negative feedback loop regulation may control the beta-catenin/Tcf pathway. This signaling also upregulated NF-kappaB transactivation without affecting the activity of IkappaB kinase, thereby establishing that the maintenance of the betaTrCP level is important for coordination between beta-catenin/Tcf and NF-kappaB signaling.
Cytoskeletal Proteins↗
K-ras mutation: early detection in molecular diagnosis and risk assessment of colorectal, pancreas, and lung cancers--a review.
Multiple genomic alterations are involved in the development of most human cancers. They include alterations in oncogenes, tumor suppressor genes, DNA mismatch repair and excision repair genes. Genetic testing for susceptibility has been a part of the management of patients with well-defined but uncommon hereditary cancers in which certain susceptible gene mutations are determined in the germ line. However, a molecular diagnostic approach to sporadic cancers, which comprise the vast majority of malignant tumors in human beings, is still under development. One of the best characterized tumor-related genes is K-ras, which somatically mutates in several types of sporadic human cancers. Since mutations of this gene occur exclusively in three hot spots (codons 12, 13 and 61), and are frequently detected and well characterized in colorectal, pancreas and lung cancers, molecular diagnosis and susceptibility (risk) assessment targeting K-ras mutations are being developed. For this purpose, sample collection methods that reflect the state of the entire affected organ are important. Clinical samples used for molecular diagnosis and risk assessment include stool and lavage fluid, pancreatic and duodenal juices, and sputum and lavage fluids for colorectal, pancreas and lung cancers, respectively. The reported incidence of K-ras mutations detected in these samples ranges from 7% to 80% for colorectal cancers, 25% to 87% for pancreatic cancers, and 25% to 48% for lung cancers. Incidence of mutations clearly depends on the sensitivity of the method for detecting the mutant K-ras allele, as well as the nature and the quality of the clinical samples. Various methods including plaque hybridization, dot blot hybridization, combined PCR and RFLP or SSCP, and sensitive PCR have been used, and they exhibited high specificity (75 to 100%) in detecting mutations. Molecular analysis is demonstrating promise in assessing susceptibility to, or risk of developing, sporadic cancers.
Biomarkers, Tumor↗
Altered expression of p53 and p27 proteins, alone or combined, as a predictor of metastatic potential in early invasive carcinoma of colon and rectum--a comparative clinicopathologic and molecular analysis.
To place the choice of therapy (endoscopic resection or radical surgery) in early invasive carcinoma (EIC) of colon and rectum on a more rational basis, this study sought to identify molecular predictors of metastasis. Several morphologic risk factors (histologic type, degree of tumor invasion, lymphatic and venous invasion) and expression of p53 and p27 proteins in the primary tumor were compared in 80 patients with EIC, including 12 (15%) with metastasis or recurrence (or both). Of the factors enumerated, deeper invasion of the submucosal layer, lymphatic-venous invasion, p53 overexpression, and decreased expression of p27 were correlated significantly with metastasis. The results also indicated that altered expression of p53 or p27 is independently relevant to metastasis of EIC. Analysis of these markers, together with determination of the morphologic risk factors, could complement the identification of patients with metastasis on the basis of known morphologic risk factors. Because the molecular factors can be assessed more objectively than can the morphologic parameters, they may strengthen the ability to identify EIC that has undergone, or will undergo, metastasis.
Adenocarcinoma↗
Altered expression of beta-catenin and c-erbB-2 in early gastric cancer.
To investigate the possible relationship between altered expression (loss of membranous staining or nuclear accumulation) of beta-catenin and invasion/metastasis in early gastric cancer (EGC), beta-catenin was detected immunohistochemically in 116 cases of EGC, including 86 differentiated and 30 undifferentiated carcinomas. In parallel, immunohistochemical expression of c-erbB-2 was analyzed in all EGC cases. Regardless of histological type, altered expression of beta-catenin was found in 47% of mucosal carcinomas and 89% of carcinomas with submucosal invasion (p<0.001). Of particular interest is that beta-catenin alteration was found in almost all EGCs with lymph node metastasis, even though no significant statistical comparison could be made. These results suggest that molecular changes resulting in abnormal beta-catenin expression participate in the process of submucosal invasion and metastasis. While loss of expression was preferentially observed in undifferentiated EGCs, nuclear accumulation was found exclusively in 24% of differentiated EGCs. c-erbB-2 was overexpressed in only 16% of differentiated EGCs but there was no correlation between this overexpression and invasion or metastasis. However, it is intriguing that 12 out of 14 cases with c-erbB-2 overexpression also showed altered beta-catenin expression, suggesting that both molecules are involved in the development of a certain set of differentiated EGCs.
Adenocarcinoma↗
Environmental factors as regulators and effectors of multistep carcinogenesis.
This review highlights current knowledge of environmental factors in carcinogenesis and their cellular targets. The hypothesis that environmental factors influence carcinogenesis is widely supported by both epidemiological and experimental studies. The fact that only a small fraction of cancers can be attributed to germline mutations in cancer-related genes further buttresses the importance of environmental factors in carcinogenesis. Furthermore, penetrance of germline mutations may be modified by either environmental or other genetic factors. Examples of environmental factors that have been associated with increased cancer risk in the human population include chemical and physical mutagens (e.g. cigarette smoke, heterocyclic amines, asbestos and UV irradiation), infection by certain viral or bacterial pathogens, and dietary non-genotoxic constituents (e.g. macro- and micronutrients). Among molecular targets of environmental influences on carcinogenesis are somatic mutation (genetic change) and aberrant DNA methylation (epigenetic change) at the genomic level and post-translational modifications at the protein level. At both levels, changes elicited affect either the stability or the activity of key regulatory proteins, including oncoproteins and tumor suppressor proteins. Together, via multiple genetic and epigenetic lesions, environmental factors modulate important changes in the pathway of cellular carcinogenesis.
Biomarkers↗
Collagen production by cultured human retinal pigment epithelial cells.
We investigated extracellular matrix produced by human retinal pigment epithelial cells (RPE) in vitro using electron microscopy and enzyme linked immunosorbent assay (ELISA). The thickness of the matrix under the cell layer was about 30 microns after 360 days of culture. It consisted mainly of fibrous and granular components. Type IV and V collagen were detected but type I and III were not detected by ELISA. It appeared that RPE can secrete type IV and V collagen and form a thick membrane which may cause proliferative vitreoretinopathy (PVR) by contraction. Control of RPE proliferation and secretion of extracellular matrix is indispensable for prevention of PVR.
Cells, Cultured↗
Conformation-dependent phosphorylation of p53.
Phosphorylation of the p53 tumor suppressor protein is known to modulate its functions. Using bacterially produced glutathione S-transferase (GST)-p53 fusion protein and baculovirus-expressed histidine-tagged p53 ((His)p53), we have determined human p53 phosphorylation by purified forms of jun-N-kinase (JNK), protein kinase A (PKA), and beta subunit of casein kinase II (CKIIbeta) as well as by kinases present in whole cell extracts (WCEs). We demonstrate that PKA is potent p53 kinase, albeit, in a conformation- and concentration-dependent manner, as concluded by comparing full-length with truncated forms of p53. We further demonstrate JNK interaction with GST-p53 and the ability of JNK to phosphorylate truncated forms of GST-p53 or full-length (His)p53. Dependence of phosphorylation on conformation of p53 is further supported by the finding that the wild-type form of p53 (p53wt) undergoes better phosphorylation by CKIIbeta and by WCE kinases than mutant forms of p53 at amino acid 249 (p53(249)) or 273 (p53(273)). Moreover, shifting the kinase reaction's temperature from 37 degrees C to 18 degrees C reduces the phosphorylation of mutant p53 to a greater extent than of p53wt. Comparing truncated forms of p53 revealed that the ability of CKIIbeta, PKA, or WCE kinases to phosphorylate p53 requires amino acids 97-155 within the DNA-binding domain region. Among three 20-aa peptides spanning this region we have identified residues 97-117 that increase p53 phosphorylation by CKIIbeta while inhibiting p53 phosphorylation by PKA or WCE kinases. The importance of this region is further supported by computer modeling studies, which demonstrated that mutant p53(249) exhibits significant changes to the conformation of p53 within amino acids 97-117. In summary, phosphorylation-related analysis of different p53 forms in vitro indicates that conformation of p53 is a key determinant in its availability as a substrate for different kinases, as for the phosphorylation pattern generated by the same kinase.
Adenosine Triphosphate↗
Quantitative enriched PCR (QEPCR), a highly sensitive method for detection of K-ras oncogene mutation.
We have developed a rapid and highly sensitive method for the detection of mutant K-ras codon 12 allele in the presence of 10(5) copies of the wild-type alleles. This sensitivity is achieved by selective amplification of mutant K-ras sequences, using a two-stage procedure with modified primers. In the first stage, primers consist of K-ras sequences in the 3' portion and polyomavirus sequence (to minimize homology with human genome) on the 5' portion. The 3' portion also consists of mismatch sequence that generates an MvaI site in normal, but not mutant, K-ras codon 12 alleles. Thus, following the first round of 20 cycles, restriction enzyme cleavage is carried out to selectively digest normal K-ras codon 12 alleles. To enrich mutant alleles, a second amplification is performed using tail primers that recognize the polyoma, but not human sequences. This design ensures that in the second amplification only mutant alleles that were pre-amplified in the first round would serve as template for this reaction. Ethidium bromide-stained polyacrylamide gel electrophoresis (PAGE) of second-stage PCR product that has been digested with MvaI is used to monitor the presence of mutant alleles, detected at sensitivity of 1/10(5). This technique offers high sensitive detection of mutant K-ras alleles using a new concept of tail-primer design and is likely to assist in identifying patients at risk to develop pancreatic, colon, or lung cancer, which harbor high incidence of mutant ras alleles.
Base Sequence↗
Sclerosing pancreatitis showing rapidly progressive changes with recurrent mass formation.
CONCLUSION: Sclerosing pancreatitis might develop repeatedly or might rapidly extend to the whole pancreas with recurrent mass formation. BACKGROUND: Nothing is known concerning course or development of sclerosing pancreatitis. METHODS: A 63-yr-old male was followed up for 2.5 yr. RESULTS: The patient was admitted because of a tumor in the body and tail of the pancreas. Serum pancreatic enzymes were transiently elevated, but tumor markers were all negative. Imaging studies showed a tumor 7 cm in size. The main pancreatic duct was normal in the head and obstructed at the body on endoscopic retrograde pancreatography (ERCP). The K-ras oncogene mutation was positive in pure pancreatic juice. Distal pancreatectomy was performed because pancreatic cancer was highly suspected. Pathological findings showed that the tumor was a densely fibrotic mass without malignant cells. Inflammatory cell infiltration was observed in the stroma. One year later, another mass 3 cm in size was noted in the remnant pancreatic head. ERCP revealed diffuse irregular narrowing of the main pancreatic duct, its branches, and the common bile duct. Liver dysfunction improved and an elevation of serum pancreatic enzymes subsided without any specific treatment, and the mass diminished in size. The patterns of various imaging studies on the second tumor were the same as those of the previous resected mass. Corticosteroid was not administered.
Genes, ras↗
Does p53 overexpression cause metastases in early invasive colorectal adenocarcinoma?
OBJECTIVE: To find out whether p53 overexpression correlates with metastatic potential and other adverse prognostic factors in early invasive colorectal carcinoma and whether measurement of the expression of p53 protein could be helpful in the choice of treatment (endoscopic/local or radical resection). DESIGN: Retrospective study. SETTING: University hospital, Japan. SUBJECTS: Overexpression of p53 protein in the primary tumour was examined immunohistochemically in 50 patients with early invasive colorectal cancer. MAIN OUTCOME MEASURES: Differences in p53 overexpression between subgroups. RESULTS: Abnormal accumulation of nuclear p53 was detected in the primary tumour of 20 patients (40%) with early invasive colorectal cancer. We found p53-positive cells in 7 (78%) of 9 that had metastasised to regional lymph nodes or distant organs, or both, and in 13 (32%) of 41 that had not metastasised (p = 0.02). p53 Immunoreactivity was also present in 10 (71%) of 14 superficial (type II) lesions compared with 10 (28%) of 36 protruding (type I) ones (p = 0.009) and in 12 (57%) of 21 moderately differentiated adenocarcinomas compared with 8 (28%) of 29 well-differentiated adenocarcinomas (p = 0.045). There was no significant correlation between p53 overexpression and the depth of tumour invasion or angiolymphatic involvement. The p53-positive metastasising tumours had features that corresponded to those of early carcinoma arising de novo. CONCLUSION: Our results seem to support the postulate that p53 overexpression in early invasive colorectal carcinomas is associated with an increase in their metastatic potential.
Adenocarcinoma↗
[Correlations between risk factors for metastasis in early tumors of the large intestine].
UNLABELLED: Intramucosal colorectal cancers (Tis) are treated by local excision. Controversies remain as to the management of patients with the submucosally invasive colorectal cancers (EIC). The decision for radical surgery or local excision depends on an estimate of the risk of metastases. AIM: To estimate correlations between the risk factors for metastases in EIC and to evaluate the tests that identify the metastasising EICs. In the study, we investigated the relationships between selected prognostic factors (the gross appearance, depth of submucosal invasion, vessel invasion, grade, presence of adenoma component, and p53 overexpression) and the frequency of metastases in 64 EICs. STATISTICAL ANALYSIS: Fisher's exact test and log-linear analysis. Tests' evaluation: sensitivity, specificity, predictive values. The metastases were detected in 11 tumours (17%). The metastases were found with the significantly (p < 0.05) higher frequency in moderately differentiated adenocarcinomas, in tumours with massive invasion, with vessel invasion, and with the p53 overexpression. The only interaction was between the grade and the depth of invasion. No other interaction between the risk factors was found with the multivariate analysis. The evaluation of the depth of invasion was characterised by the highest sensitivity in detecting metastases, and by the highest negative predictive value. The highest specificity and the positive predictive value were found for the evaluation of vessel invasion. The massive submucosal invasion, vessel invasion, moderately differentiated characteristics, and the p53 overexpression seem to be the independent risk factors for metastases in EIC. The evaluation of these factors should be helpful as prognostic tests and for the proper choice of the treatment modality for the patients with EIC.
Adenocarcinoma↗
Combined analysis of microsatellite instability and K-ras mutation increases detection incidence of normal samples from colorectal cancer patients.
Microsatellite instability (MI) and K-ras oncogene mutation have been widely used as biomarkers of genetic changes in colorectal cancer (CRC). Each of these biomarkers was independently found in normal-appearing colonic mucosa at stages preceding the development of CRC, albeit at a relatively low incidence. To assess the potential value of combined MI and K-ras mutation analysis in the detection of normal-appearing colonic mucosa samples taken from patients with CRC, we have chosen to analyze multiple (3-7) normal colonic mucosa samples and the respective colorectal tumor tissues from 20 patients with CRC. As a control, we have used 54 normal mucosa samples obtained from 9 autopsies of patients without CRC. In at least 1 of 5 loci analyzed, MI was found in 8 of 20 patients via analysis of multiple normal-appearing colonic mucosa samples from each patient. Combined analysis of MI and mutant ras alleles in normal-appearing colonic mucosa samples enabled the identification of 11 of 20 patients with CRC. None of the 54 normal colonic mucosa samples obtained from 9 patients without CRC were found to carry mutant ras or MI. The ability to detect 55% of patients with CRC via the analysis of normal mucosa samples provides an important advance in our approach toward early detection of individuals who may be at risk to develop this tumor type.
Aged↗