ABSTRACT
Nager syndrome belongs to the group of acrofacial dysostosis, which are characterized by the association of craniofacial and limb malformations. Recently, exome sequencing studies identified the SF3B4 gene as the cause of this condition in most patients. SF3B4 encodes a highly conserved protein implicated in mRNA splicing and bone morphogenic protein (BMP) signaling. We performed SF3B4 sequencing in 14 families (18 patients) whose features were suggestive of Nager syndrome and found nine mutations predicted to result in loss-of-function. SF3B4 is the major gene responsible for autosomal dominant Nager syndrome. All mutations reported predict null alleles, therefore precluding genotype-phenotype correlations. Most mutation-negative patients were phenotypically indistinguishable from patients with mutations, suggesting genetic heterogeneity.
Subject(s)
Genetic Predisposition to Disease/genetics , Haploinsufficiency/genetics , Mandibulofacial Dysostosis/genetics , Phenotype , RNA-Binding Proteins/genetics , Base Sequence , Female , Genes, Dominant/genetics , Humans , Male , Mandibulofacial Dysostosis/pathology , Molecular Sequence Data , Mutation/genetics , RNA Splicing Factors , Sequence Analysis, DNAABSTRACT
Split hand/foot malformation (SHFM) with long-bone deficiency (SHFLD, MIM#119100) is a rare condition characterized by SHFM associated with long-bone malformation usually involving the tibia. Previous published data reported several unrelated patients with 17p13.3 duplication and SHFLD. Recently, the minimal critical region had been reduced, suggesting that BHLHA9 copy number gains are associated with this limb defect. Here, we report on 13 new families presenting with ectrodactyly and harboring a BHLHA9 duplication.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/genetics , Genes, Duplicate , Limb Deformities, Congenital/genetics , Tibia/abnormalities , Chromosomes, Human, Pair 17/genetics , Female , Humans , Limb Deformities, Congenital/physiopathology , Male , Pedigree , Phenotype , Tibia/physiopathologyABSTRACT
BACKGROUND: Microsatellite instability (MSI) is a molecular phenotype due to defective DNA mismatch repair (MMR) system. It is used to predict outcome of colorectal tumours and to screen tumours for Lynch syndrome (LS). A pentaplex panel composed of five mononucleotide markers has been largely recommended for determination of the MSI status. However, its sensitivity may be taken in default in occasional situations. The aim of the study was to optimise this panel for the detection of MSI. METHODS: We developed an assay allowing co-amplification of six mononucleotide repeat markers (BAT25, BAT26, BAT40, NR21, NR22, NR27) and one polymorphic dinucleotide marker (D3S1260) in a single reaction. Performances of the new panel were evaluated on a cohort of patients suspected of LS. RESULTS: We demonstrate that our assay is technically as easy to use as the pentaplex assay. The hexaplex panel shows similar performances for the identification of colorectal and non-MSH6-deficient tumours. On the other hand, the hexaplex panel has higher sensitivity for the identification of MSH6-deficient tumours (94.7% vs 84.2%) and MMR-deficient tumours other than colorectal cancer (92.9% vs 85.7%). CONCLUSION: The hexaplex panel could thus be an attractive alternative to the pentaplex panel for the identification of patients with LS.
Subject(s)
Biomarkers, Tumor , DNA Mismatch Repair/genetics , Early Detection of Cancer/methods , Microsatellite Repeats , Neoplasms/diagnosis , Adult , Biomarkers, Tumor/analysis , Biomarkers, Tumor/genetics , Case-Control Studies , DNA Repair-Deficiency Disorders/diagnosis , DNA Repair-Deficiency Disorders/genetics , Female , Fluorescence , Genes, Neoplasm , Humans , Microsatellite Instability , Microsatellite Repeats/physiology , Middle Aged , Neoplasms/genetics , Polymerase Chain Reaction/methodsABSTRACT
BACKGROUND: Activator protein-2alpha (AP-2alpha) is a transcription factor that belongs to the family of AP-2 proteins that have essential roles in tumorigenesis. Indeed, AP-2alpha is considered as a tumour-suppressor gene in different tissues such as colonic, prostatic or breast epithelial cells. Moreover, AP-2alpha also participates in the control of colon and breast cancer cells sensitivity towards chemotherapeutic drugs. Despite its potential interest, very few data are available regarding the roles of AP-2alpha in pancreatic cancer. METHODS: We have developed a stable pancreatic CAPAN-1 cell line overexpressing AP-2alpha. Consequences of overexpression were studied in terms of in vivo cell growth, gene expression, migration capacity and chemosensitivity. RESULTS: In vivo tumour growth of CAPAN-1 cells overexpressing AP-2alpha was significantly decreased by comparison to control cells. An altered expression pattern of cell cycle-controlling factors (CDK-4, CDK-6, cyclin-G1, p27(kip1) and p57(kip2)) was observed in AP-2alpha-overexpressing clones by microarrays and western blot analysis. Promoter activity and ChIP analysis indicated that AP-2alpha induces p27(kip1) protein levels by direct binding to and transactivation of its promoter. Moreover, AP-2alpha overexpression increased the chemosensitivity of CAPAN-1 cells to low doses of gemcitabine and reduced their in vitro migration capacity. CONCLUSION: Our data suggested that AP-2alpha overexpression could be exploited to decrease in vivo tumour growth of pancreatic cancer cells and to increase their sensitivity to gemcitabine.
Subject(s)
Antimetabolites, Antineoplastic/pharmacology , Deoxycytidine/analogs & derivatives , Drug Resistance, Neoplasm/genetics , Pancreatic Neoplasms/genetics , Transcription Factor AP-2/genetics , Animals , Blotting, Western , Cell Line, Tumor , Cell Movement , Cell Proliferation/drug effects , Deoxycytidine/pharmacology , Gene Expression , Humans , Mice , Mice, Nude , Microscopy, Confocal , Oligonucleotide Array Sequence Analysis , Pancreatic Neoplasms/metabolism , Transcription Factor AP-2/metabolism , Transfection , Xenograft Model Antitumor Assays , GemcitabineABSTRACT
Calcium is a major ion in human metabolism and its level is highly controlled. This regulation is performed via the Calcium Sensing Receptor, a discovery which ten years ago led to the explanation of a number of clinical disorders. The syndromes caused by CaSR abnormalities are characterized by hypercalcemia or hypocalcemia, associated with inappropriate calciuria. An underlying genetic or auto-immune cause may be demonstrated. High blood calcium levels linked to mutations of the CaSR gene lead to familial hypocalciuric hypercalcemia and the neonatal and non neonatal forms with severe hypercalcemic. Hypocalcemia determined by mutations in the CaSR gene include autosomal dominant hypocalcemia and its sporadic form. Another clinical presentation similar to Bartter syndrome has been reported. Auto-antibodies directed against CaSRs, seen in auto-immune diseases, can lead to similar clinical presentations. Finally, CaSR polymorphisms modulate the range of blood calcium levels. With diagnosis of these diseases deleterious therapeutics can be avoided. The discovery of this receptor has led to new therapeutic prospects such as calcimimetics for hyperthyroidism.
Subject(s)
Receptors, Calcium-Sensing/genetics , Chromosome Mapping , Chromosomes, Human, Pair 3 , Humans , Hypercalcemia/genetics , Hypercalcemia/pathology , Hypocalcemia/pathology , Infant, Newborn , MutationABSTRACT
Multiple Endocrine Neoplasia type 1 (MEN1) is an autosomal dominant hereditary syndrome (OMIM 131100) due to MEN1 gene mutations, predisposing to the development of hyperplasic and tumoral lesions of neuroendocrine tissues. Since the identification of the gene in 1997, more than 400 different mutations of MEN1 have been registered. Genotypic analysis of MEN1 remains fastidious and must be reserved to targeted situations. If the lesions appear in a familial assessed context, there is a strong argument to search for MEN1 mutation. This is not the case in a sporadic context. With experience acquired in our laboratory, we evaluated the frequency of MEN1 mutations in patients with sporadic presentations. Our aim was to better define criteria for MEN1 genotypic analysis. One hundred and twenty four blood samples from unrelated patients, who gave their written informed consent, were analyzed. These patients exhibited 1 to 4 manifestations of MEN1 without any familial context. After DNA extraction, the analysis was undertaken by PCR-sequencing of all the MEN1 coding exons and exon/intron boundaries or by PCR of the pre-screened fragments alone, a technique made possible by indirect screening mutation methods. Mutations were identified by comparing the sequences to the reference MEN1 sequence available from GENBANK (U93237.1). Mutations were identified in 19 patients, with variable prevalence according to clinical manifestations: 100% for patients with 4 manifestations, 45.5% for patients with 3 manifestations, 19% for patients with 2 manifestations and 2% for patients with only one manifestation. Mutations were: 11 point variations (58%), including 2 splicing sites and 8 frameshift mutations (42%) including 5 deletions, 2 insertions and 1 insertion/deletion; one mutation was identified twice. We showed a relationship between clinical presentation and MEN1 mutation identification, especially with the number of clinical manifestations but also with the type of manifestation. Pancreatic manifestations were significantly linked with probability of mutation. In a sporadic context with at least two established manifestations of MEN1, the overall probability of identifying a mutation was 26%, warranting MEN1 genotypic analysis.
Subject(s)
Chromosomes, Human, Pair 11 , Genetic Testing , Multiple Endocrine Neoplasia Type 1/genetics , Adult , DNA/blood , DNA/genetics , DNA/isolation & purification , Diagnosis, Differential , Gene Frequency , Humans , Middle Aged , Molecular Sequence Data , Multiple Endocrine Neoplasia Type 1/classification , Multiple Endocrine Neoplasia Type 1/diagnosis , MutationABSTRACT
The amino acid sequence of the proline-rich glycoprotein from human parotid saliva was studied using different preparations: native glycoprotein, tryptic glycopeptides, and their chemically deglycosylated homologous derivatives. The results indicate very similar structural features of the proline-rich glycoprotein and the tryptic glycopeptides and suggest that the peptide backbone of the glycoprotein comprises several repetitive domains containing one carbohydrate-peptide linkage and a proline-rich sequence. These data are in fair agreement with a preceding circular dichroism study (Aubert, J.P., Porchet, N., Boersma, A., Loucheux-Lefevbre, M.H. and Degand, P. (1982) Biochim. Biophys. Acta 704, 361-365).
Subject(s)
Glycoproteins/analysis , Parotid Gland/metabolism , Proline/analysis , Saliva/analysis , Amino Acid Sequence , Carbohydrates/analysis , Chromatography, Gel , Humans , Peptide Fragments/analysis , Trypsin/analysisABSTRACT
Previous results on the primary structure of the human parotid saliva proline-rich glycoprotein established the concept of repetitive domains in the sequence of that glycoprotein, tryptic glycopeptides of proline-rich glycoprotein representing the basic structure. The present work is concerned with the study of the secondary structure of the proline-rich glycoprotein, and of tryptic glycopeptides with and without the glycan moiety, using circular dichroism. CD spectra exhibit the same secondary structure with about 60% of polyproline II helical structure for the proline-rich glycoprotein, tryptic glycopeptides and their deglycosylated homologues. The present results are in fair agreement with the amino acid sequence results and suggest a model for the schematic representation of the proline-rich glycoprotein.
Subject(s)
Glycoproteins , Salivary Proteins and Peptides , Calcium , Circular Dichroism , Humans , Parotid Gland , Peptide Fragments , Proline , Protein ConformationABSTRACT
THIS RETROSPECTIVE STUDY AIMS: To define a clinical and secretory profile of paragangliomas extra-adrenal chromaffin tumors. METHODS: From 1971 throughout 2002, 39 paragangliomas have been observed in 38 patients (22 male, 16 female, average age 41,2 years). RESULTS: Four were located above the diaphragm, 35 were sub-phrenic (6 of the organ of Zuckerkandl), 32 secreted catecholamines, 23 were hypertensive (with only one without hypersecretion of catecholamines). Among 29 (131)I-metaiodobenzylguanidine scans (MIBG) reviewed, 20 tumors took up the radiopharmaceutical. The treatment was surgical in 35 cases with addition of external radiotherapy and MIBG in one case each; two patients died before any treatment. Two patients with persistent disease after surgery were successfully treated by surgery or MIBG. Histologically, 20 were malignant and 17 were seemingly benign. All exclusive dopamine secreting paragangliomas were malignant. Six patients relapsed two of which for a tumor initially classified as benign. The treatment of recurrences was surgical, by MIBG or by external radiotherapy. Nine patients had a family history of chromaffin tumor(s). The genetic survey made in five of these nine patients was positive in all cases.
Subject(s)
Adrenal Gland Neoplasms/metabolism , Adrenal Gland Neoplasms/pathology , Catecholamines/metabolism , Paraganglioma/metabolism , Paraganglioma/pathology , 3-Iodobenzylguanidine/therapeutic use , Adrenal Gland Neoplasms/genetics , Adrenal Gland Neoplasms/therapy , Adult , Antineoplastic Agents/therapeutic use , Female , Humans , Male , Paraganglioma/genetics , Paraganglioma/therapy , Retrospective Studies , Treatment OutcomeABSTRACT
Familial medullary thyroid carcinoma (FMTC) and multiple endocrine neoplasia type 2A syndromes are dominantly inherited diseases caused by activating germline mutations of the RET protooncogene. The majority of these patients carry a germline point mutation affecting one of five cysteine residues encoded by exon 10 (codon 609, 611, 618, or 620) or 11 (codon 634). In a few FMTC families, point mutations involving noncysteine codons in exon 13 (codons 768, 790, and 791), 14 (codon 804), or 15 (codon 891) have been reported. Hirschsprung's disease is a nonneoplastic disorder associated with RET mutations leading to a loss of function effect. Mutations are identified in 50% of the familial cases and are scattered along the gene. We now report the study of a FMTC family with four affected members and a history of fatal neonatal intestinal obstruction in the sister of the proband. Genetic analysis demonstrated the absence of an usual FMTC mutation and the presence of a germline 9-bp duplication in RET exon 8 in the heterozygous state in all patients with MTC. This new mutation creates an additional cysteine residue in the extracellular cysteine-rich domain of RET. Further studies are warranted to confirm whether this new mutation is causing MTC only or could be associated with Hirschsprung's disease.
Subject(s)
Carcinoma, Medullary/genetics , Exons , Membrane Proteins/genetics , Thyroid Neoplasms/genetics , Adult , Child , Coatomer Protein , Cysteine/blood , Cysteine/genetics , Female , Germ-Line Mutation , Hirschsprung Disease/genetics , Humans , Male , Middle Aged , Mutation , Pedigree , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The cells of living organisms in contact with the external environment are constantly attacked by different kinds of substances such as micro-organisms, toxins, and pollutants. With evolution, defense mechanisms, such as the secretion of mucus has been developed. Mucins are the main components of mucus. They are synthesized and secreted by specialized cells of the epithelium and in some case, by non mucin-secreting cells. Little was known about the structure of mucins until a decade ago. This is principally due to heavy glycosylation of mucins, which complicated their analysis. With the application of molecular biological methods, structures of the mucin core peptides (apomucins) are beginning to be elucidated. A total of eleven human mucin (MUC) genes have been identified and numbered in chronological order of their description: MUC1-4, MUC5AC, MUC5B, MUC6-8, and MUC11-12. Of these, the complete cDNA sequence are published only for six mucins MUC1, MUC2, MUC4, MUC5B, MUC5AC, and MUC7. Human mucin genes, in general, show three common features: I) a nucleotide tandem repeat domain; II) a predicted peptide domain containing a high percentage of serines and threonines; III) complex RNA expression. The tandem repeats in mucins make up the majority of the backbone. Related to their structure, mucins can be classified in three distinct sub-families: gel-forming, soluble, and membrane-bound. Each member from one family possesses common characteristics and probably specific functions. For a long time, they were thought to have the unique function of protecting and lubricating the epithelial surfaces. The study of the mucins structure as well as the relationship between structure and function show that mucins also possess other important functions, such as growth, direct implication in the fetal development, the epithelial renewal and differentiation, the epithelial integrity, carcinogenesis, and metastasis. This review presents the actual knowledge on the mucins structure and the best-characterized function related to their structure.
Subject(s)
Genes/genetics , Mucins/genetics , Humans , Mucins/classification , Protein Isoforms/geneticsABSTRACT
Mucin production and secretion by specialized epithelial cells is a common mechanism used by mammals to protect the underlying mucosae against various injuries (pollutants, pathogens, pH). The expression of mucin genes is cell- and tissue-specific but is submitted to variations during cell differentiation, inflammatory process, and is altered during carcinogenesis. The molecular mechanisms responsible for the control of mucin transcription and expression are beginning to be understood as mucin gene promoters and regulatory regions are characterized. The four gel-forming mucin genes, MUC2-MUC5AC-MUC5B-MUC6, are clustered on the p15 arm of chromosome 11. Common regulatory mechanisms (PKA, PKC, PKG and Ca2+ signaling, Sp1/Sp3) may account for the capability of mucous-secreting cells to express several mucin genes simultaneously. In response to an insult or during carcinogenesis, the normal pattern of expression is altered and results from specific answers of the cell by activating different intracellular signaling pathways. 11p15 mucin genes are regulated at the transcriptional level by pro-inflammatory cytokines (IL-1beta, IL-6, TNF-alpha), pleiotropic cytokines (IL-4, IL-13, IL-9), bacterial exoproduct (LPS), growth factors (EGF, TGF-alpha), lipid mediator (PAF), retinoids and hormones. To date, the only downstream cascade known to activate mucin gene transcription is the Src/Ras/MAPK/pp90rsk cascade, which leads to the activation of the transcription factor NF-kappaB. Mucin gene transcription is also regulated by ATF-1, CREB and RAR-alpha transcription factors. Finally, repression of mucin transcription in cancer cells is under the control of the epigenetic mechanism of methylation. As transcriptional regulation of mucin genes begins to be unraveled, it becomes clear that many signaling pathways are involved. Our understanding of mucin gene transcriptional regulation, which awaits more data (identification of the signaling cascades and active cis-elements within promoters and introns), will most certainly lead to the use of mucin genes as molecular markers in cancer and molecular tools in human gene therapy, and to the synthesis of new therapeutic agents in inflammatory diseases of the epithelium.
Subject(s)
Chromosomes, Human, Pair 11/genetics , Gene Expression Regulation , Inflammation/genetics , Mucins/genetics , Multigene Family , Neoplasms/genetics , Humans , Inflammation/pathology , Multigene Family/genetics , Neoplasms/pathology , Promoter Regions, Genetic/genetics , Transcription, GeneticABSTRACT
Mucins are glycoproteins synthesized by epithelial cells and thought to promote tumor-cell invasion. Eight human mucin genes have been well characterized: MUC2, MUC5AC, MUC5B, MUC6 map to 11p15.5 and encode secretory gel forming mucins while MUC1, MUC3, MUC4, MUC7 are scattered on different chromosomes and encode membrane-bound or secreted mucins. The expression pattern of the mucin genes is complex in normal airways involving six genes, mainly MUC5AC and MUC5B in mucus-producing cells and MUC4 in a wide array of epithelial cells. MUC5AC overexpression in metaplasia, dysplasia and normal epithelium adjacent to squamous cell carcinoma provides additional arguments for a mucous cell origin of preneoplastic squamous lesions. MUC5AC and MUC5B expression is related to mucus formation in adenocarcinomas. Mucinous bronchioloalveolar carcinoma (BAC) has a particular pattern of mucin gene expression indicating that it has sustained a well-differentiated phenotype similar to the goblet cell, correlated with distinctive features i.e. a noninvasive pattern and a better prognosis than nonBACs. MUC4 is the earlier mucin gene expressed in the foregut, before epithelial differentiation and is expressed independently of mucus secretion both in normal adult airways and carcinomas. These findings are in favor the histogenetic theory of non-small-cell carcinoma originating from a pluripotent mucous cell.
Subject(s)
Lung Neoplasms/genetics , Mucins/genetics , Precancerous Conditions/genetics , Respiratory Mucosa/metabolism , Gene Expression Regulation , Humans , Protein Isoforms/geneticsABSTRACT
In recent years considerable advances have been made in our knowledge of the peptide moiety of human mucins through cDNA cloning. In many diseases disorders in mucin biosynthesis are observed, which result either from changes in the synthesis of the carbohydrate side chains or from differences in the relative expression of the different apomucins, each of which may affect physical properties of the viscous gel. We describe in situ hybridization studies on healthy human mucosae with five different oligonucleotide probes corresponding to each of the human genes known to date that encode secreted mucins, i.e., MUC 2, 3, 4 (HGM nomenclature) and 5B, 5C (proposed name). These genes present a nucleic tandem repeat organization. The choice of oligonucleotide probes was made to amplify the signal by hybridization of many small probes on the same mRNA molecules. A characteristic pattern of mucin gene expression was observed for each mucosa.
Subject(s)
Cervix Uteri/chemistry , Digestive System/chemistry , Gene Expression , Lung/chemistry , Mucins/genetics , Base Sequence , Blotting, Northern , Bronchi/chemistry , Epithelium/chemistry , Female , Gastric Mucosa/chemistry , Humans , In Situ Hybridization , Intestines/chemistry , Molecular Sequence Data , Mucous Membrane/chemistry , Oligonucleotide Probes , RNA, Messenger/analysisABSTRACT
Studies were undertaken to provide information regarding cell-specific expression of mucin genes in stomach and their relation to developmental and neoplastic patterns of epithelial cytodifferentiation. In situ hybridization was used to study mRNA expression of eight mucin genes (MUC1-4, MUC5AC, MUC5B, MUC6, MUC7) in stomach of 13 human embryos and fetuses (8-27 weeks' gestation), comparing these with normal, metaplastic, and neoplastic adult tissues. These investigations have demonstrated that MUC1, MUC4, MUC5AC, MUC5B, and MUC6 are already expressed in the embryonic stomach at 8 weeks of gestation. MUC3 mRNA expression can be observed from 10.5 weeks of gestation. MUC2 is expressed at later stages, concomitant with mucous gland cytodifferentiation. Normal adult stomach is characterized by strong expression of MUC1, MUC5AC, and MUC6, less prominent MUC2, and sporadic MUC3 and MUC4, without MUC5B and MUC7. Intestinal metaplasia is characterized by an intestinal-type pattern with MUC2 and MUC3 mRNA expression. Gastric carcinomas exhibit altered mucin gene expression patterns with disappearance of MUC5AC and MUC6 mRNAs in some tumor glands, abnormal expression of MUC2, and reappearance of MUC5B mRNAs. In conclusion, we have observed that patterns of mucin gene expression in embryonic and fetal stomach could show similarities with some gastric carcinomas in adults. Differences in mucin gene expression in developmental, metaplastic, and neoplastic stomach compared to normal adult stomach suggest a possible regulatory role for their products in gastric epithelial cell proliferation and differentiation.
Subject(s)
Adenocarcinoma/metabolism , Gastric Mucosa/metabolism , Mucins/metabolism , Stomach Neoplasms/metabolism , Adult , Gestational Age , Humans , In Situ Hybridization , Metaplasia , Mucins/genetics , RNA, Messenger/metabolism , Stomach/embryology , Stomach/pathologyABSTRACT
Studies were undertaken to provide information regarding cell-specific expression of mucin genes and their relation to developmental and neoplastic patterns of epithelial cytodifferentiation. In situ hybridization was used to study mRNA expression of mucin genes in duodenum and accessory digestive glands (liver, gallbladder, pancreas) of 13 human embryos and fetuses (6. 5-27 weeks' gestation), comparing these with normal and neoplastic adult tissues. These investigations demonstrated that the pattern of mucin gene expression in fetal duodenum reiterated the patterns we observed during gastric and intestinal ontogenesis, with MUC2 and MUC3 expression in the surface epithelium and MUC6 expression associated with the development of Brünner's glands. In embryonic liver, MUC3 was already expressed at 6.5 weeks of gestation in hepatoblasts. As in adults, MUC1, MUC2, MUC3, MUC5AC, MUC5B, and MUC6 were expressed in fetal gallbladder, whereas MUC4 was not. In contrast, MUC4 was strongly expressed in gallbladder adenocarcinomas. MUC5B and MUC6 were expressed in fetal pancreas, from 12 weeks and 26 weeks of gestation, respectively. Surprisingly, MUC3 which is strongly expressed in adult pancreas, was not detected in developmental pancreas. Taken together, these data show complex spatio-temporal regulation of the mucin genes and suggest a possible regulatory role for mucin gene products in gastroduodenal epithelial cell differentiation.
Subject(s)
Duodenum/metabolism , Gallbladder/metabolism , Liver/metabolism , Mucins/metabolism , Pancreas/metabolism , Adenocarcinoma/metabolism , Adult , Biliary Tract/metabolism , Duodenum/embryology , Gallbladder Neoplasms/metabolism , Gestational Age , Humans , In Situ Hybridization , Mucins/genetics , Organ Specificity , RNA, Messenger/metabolismABSTRACT
Our goal was to determine the effect of transdermal nicotine on cytokine and mucin gene transcription in ulcerative colitis (UC). Sixty-four nonsmoking patients with active UC were randomly assigned to transdermal nicotine (maximum dose 22 mg/day) or placebo for 4 weeks. Clinical assessment and colonic mucosal biopsies were obtained at entry and after 4 weeks. Inflammatory and immunoregulatory cytokines were assessed by qualitative reverse transcriptase-polymerase chain reaction (RT-PCR). Based on this initial screen. IL-8 mRNA levels were measured by RT-competitive PCR. MUC1, MUC2, MUC3, MUC4, MUC5AC, MUC5B, and MUC6 mRNA concentrations were measured by quantitative dot blot analysis. Cytokine mRNA expression, except for IL-8, was similar in all patients. IL-8 mRNA levels were significantly decreased in the colonic mucosa of nicotine-treated patients who improved (p = 0.04). IL-8 mRNA values were similar before and after treatment in nonresponding nicotine-treated patients and in all placebo-treated patients. Mucin gene expression was similar in all patient groups. Beneficial effects of transdermal nicotine in active UC may result from decrease of IL-8 expression at the transcriptional level. Transdermal nicotine has no effect on mucin gene transcription.
Subject(s)
Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/metabolism , Interleukin-8/biosynthesis , Mucins/biosynthesis , Nicotine/pharmacology , Administration, Cutaneous , Adult , Blotting, Northern , Colon/metabolism , Cytokines/biosynthesis , Cytokines/genetics , Double-Blind Method , Female , Gene Expression/drug effects , Humans , Interleukin-8/genetics , Intestinal Mucosa/metabolism , Male , Mucins/genetics , Nicotine/therapeutic use , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Alterations in the structure and/or quantity of mucins could alter the barrier function of mucus and play a role in initiating and maintaining mucosal inflammation in Crohn's disease. To investigate the hypothesis of a mucin gene defect in Crohn's disease, we analyzed the expression of the different mucin genes in the ileal mucosa of patients with Crohn's disease and controls. mRNA expression levels were assessed by a quantitative dot blot analysis and compared (i) between healthy and involved ileal mucosa of patients with Crohn's disease and (ii) between healthy mucosa of patients with Crohn's disease and controls. Expression of the different mucin genes was heterogeneous among controls and patients with Crohn's disease, except for MUC6 in controls. Nevertheless, MUC1 mRNA expression was significantly decreased in the involved ileal mucosa of patients with Crohn's disease when compared to the healthy mucosa (p = 0.02). Moreover, the expression levels of MUC3, MUC4, and MUC5B were significantly lower in both healthy and involved ileal mucosa of patients with Crohn's disease compared to controls (p < or = 0.05). The decrease of expression levels of some mucin genes (more particularly MUC3, MUC4, and MUC5B) in both healthy and involved ileal mucosa suggests a primary or very early mucosal defect of these genes in CD.
Subject(s)
Crohn Disease/genetics , Gene Expression Regulation , Mucins/genetics , Adolescent , Adult , Aged , Biopsy, Needle , Humans , Ileum/chemistry , Ileum/pathology , Intestinal Mucosa/chemistry , Intestinal Mucosa/pathology , Male , Middle Aged , RNA, Messenger/analysis , Reference Values , Sensitivity and Specificity , Statistics, NonparametricABSTRACT
Lung adenocarcinomas are heterogeneous clinically and histologically. Expression of the mucin genes was analyzed as a molecular marker of glandular cytodifferentiation in primary lung adenocarcinomas. Expression was correlated with histopathologic subtypes of World Health Organization classification with the aim of investigating the histogenesis of primary lung adenocarcinomas. Thirty-four primary lung adenocarcinomas were examined by in situ hybridization for mucin gene expression (MUC1-4, MUC5AC, MUC5B, MUC6-7) and by immunohistochemistry for MUC5AC and MUC5B apomucin expression. Mucinous bronchioloalveolar carcinoma (BAC) had a homogeneous pattern of mucin gene expression different from those of other types of lung adenocarcinoma, involving secreted mucins (MUC5AC, MUC5B, and MUC6) and membrane-bound mucins (MUC1, MUC3, and MUC4). Non-BAC adenocarcinoma and mucinous BAC aberrantly expressed mucin genes MUC3, and MUC3 and MUC6, respectively, which are undetectable in normal fetal and adult lung. Our results show the particular phenotype of mucin gene expression in mucinous type of BACs and the heterogeneous expression of respiratory and nonrespiratory mucins in the other types. This finding supports the theory of a common progenitor cell with the potential of multicellular differentiation. From a practical point of view, the aberrant expression of MUC3 and MUC6 could serve as a diagnostic marker in the management of the mucinous type of bronchioloalveolar carcinomas. HUM PATHOL 32:274-281.
Subject(s)
Adenocarcinoma, Bronchiolo-Alveolar/chemistry , Adenocarcinoma/chemistry , Gene Expression , Lung Neoplasms/chemistry , Mucins/genetics , Adenocarcinoma/pathology , Adenocarcinoma, Bronchiolo-Alveolar/pathology , Adult , Aged , Female , Histocytochemistry , Humans , Immunohistochemistry , In Situ Hybridization , Keratins/analysis , Lung Neoplasms/pathology , Male , Middle Aged , RNA, Messenger/analysisABSTRACT
Pulmonary maturity of the fetus can be evaluated by the lecithin/sphingomyelin (L/S) ratio in amniotic fluid. To existing methods of lipid extraction, precipitation with acetone and chromatography, we add a simple and accurate estimation of sphingomyelins (S) and precipitated lecithins (Lp) without acid digestion. The method is reproducible (C.V. less than 9%) for the measurement of Lp/S ratio and gives with accuracy the concentrations of Lp, avoiding possible errors in interpretation of Lp/S. Our results show that at 35 weeks of normal gestation, Lp/S ratio is about 2 and Lp concentration, 10 mg/1.