Adult ; Female ; Humans ; Keratin-1 ; genetics ; Keratin-9 ; genetics ; Keratoderma, Palmoplantar ; genetics ; Male ; Middle Aged ; Pedigree

In this article we reviewed the current researches on the molecular basis of epidermolytic palmoplantar keratoderma (EPPK) and the structure and function of the keratins with mutations that can cause inherited keratin disorders. Also summarized are seventeen mutations of keratin 9 in EPPK in different ethnic populations.
Humans ; Keratin-9 ; genetics ; physiology ; Keratoderma, Palmoplantar, Epidermolytic ; genetics ; pathology ; physiopathology ; Mutation

OBJECTIVETo map and identify the disease gene for the epidermolytic palmoplantar keratoderma (EPPK) in a Uighur family of China.

METHODSBlood samples were collected and genomic DNA was extracted from 48 members of the Xinjiang Uighur family. Six microsatellite repeat sequences on chromosome region 17q12-q21 and 12q13 were selected based on the two known candidate genes KRT9 and KRT1. Two-point linkage analysis and haplotype analysis were performed. Exons and their flanking intronic sequence of the KRT9 gene were amplified by polymerase chain reaction (PCR) and sequenced.

RESULTSData from the marker D17S1787 suggested linkage and yielded a Lod score of 8.65 at theta=0 by using MLINK software. Genotypes and haplotypes were acquired. The disease gene of the EPPK family is located between markers 17/TG/36620115 and D17S846. Chromosome 12q13 region was excluded with the negative Lod score obtained in marker D12S96 (Lod=-infinity at theta=0). No pathogenic mutation was detected in the KRT9 gene.

CONCLUSIONThe disease gene of the EPPK family is located on chromosome region 17q21.2. The keratin 9 gene might not be the disease gene.

China ; Chromosomes, Human, Pair 17 ; genetics ; Female ; Humans ; Keratin-1 ; genetics ; Keratin-9 ; genetics ; Keratoderma, Palmoplantar, Epidermolytic ; ethnology ; genetics ; Male ; Microsatellite Repeats ; Mutation ; Pedigree

PURPOSE: Breast cancer continues to be a major cause of death, despite the advances in the study of many prognostic factors. Although many prognostic factors have been studied, none reliably predict the response to treatment. This uncertainty in the prognostic factors could be overcome by defining the changes, occurring in patients at either the gene or protein level. Herein, attempts were made to examine the protein repertoire of patients using Proteomics. MATERIALS AND METHODS: Using conventional Proteomics, the high resolution 2-D electrophoresis followed by computational image analysis(Melanie program) and protein identification with mass spectrometry (MALDI-TOF), the serum of locally advanced breast cancer patients (stage III) was analyzed, and attempts were made to define the differences between recurred (or metastasis) patients ,and disease free patients of more than 4-years duration after surgery. RESULTS: In the 2-D electrophoresis of serum, about 1,000 spots were gained in each gel, with the up and down expressed protein spots compared to the normal control protein map. Six of seven patients had Cytokeratin 9 in their peripheral blood. In the serum of recurred patients (one of two), no Haptoglobin-related proteins were detected. All five un-recurred patients had normal or elevated levl of serum Haptoglobin-related proteins. CONCLUSIONS: The reduction of Haptoglobin-related proteins indicated the humoral immuno-depression in recurred patients. These findings may suggest the continuation of proper humoral immunity was important in the prevention of cancer recurrences or metastasis after surgery, especially in locally advanced breast cancer patients, which may suggests the value of immunotherapy in breast cancer patients to obtain good results.
Breast Neoplasms* ; Breast* ; Cause of Death ; Electrophoresis ; Humans ; Immunity, Humoral ; Immunotherapy ; Keratin-9 ; Mass Spectrometry ; Neoplasm Metastasis ; Proteomics* ; Recurrence ; Uncertainty

PURPOSE: Breast cancer continues to be a major cause of death, despite the advances in the study of many prognostic factors. Although many prognostic factors have been studied, none reliably predict the response to treatment. This uncertainty in the prognostic factors could be overcome by defining the changes, occurring in patients at either the gene or protein level. Herein, attempts were made to examine the protein repertoire of patients using Proteomics. MATERIALS AND METHODS: Using conventional Proteomics, the high resolution 2-D electrophoresis followed by computational image analysis(Melanie program) and protein identification with mass spectrometry (MALDI-TOF), the serum of locally advanced breast cancer patients (stage III) was analyzed, and attempts were made to define the differences between recurred (or metastasis) patients ,and disease free patients of more than 4-years duration after surgery. RESULTS: In the 2-D electrophoresis of serum, about 1,000 spots were gained in each gel, with the up and down expressed protein spots compared to the normal control protein map. Six of seven patients had Cytokeratin 9 in their peripheral blood. In the serum of recurred patients (one of two), no Haptoglobin-related proteins were detected. All five un-recurred patients had normal or elevated levl of serum Haptoglobin-related proteins. CONCLUSIONS: The reduction of Haptoglobin-related proteins indicated the humoral immuno-depression in recurred patients. These findings may suggest the continuation of proper humoral immunity was important in the prevention of cancer recurrences or metastasis after surgery, especially in locally advanced breast cancer patients, which may suggests the value of immunotherapy in breast cancer patients to obtain good results.
Breast Neoplasms* ; Breast* ; Cause of Death ; Electrophoresis ; Humans ; Immunity, Humoral ; Immunotherapy ; Keratin-9 ; Mass Spectrometry ; Neoplasm Metastasis ; Proteomics* ; Recurrence ; Uncertainty

OBJECTIVETo confirm that PCR products with heterozygous mutations contain not only wide-type and mutant homoduplexes, but also two types of heteroduplexes.

METHODSAn insertion-deletion mutation in the exon 1 of KRT9 gene (497delAinsGGCT), which caused Chinese epidermolytic palmoplantar keratoderma (EPPK) was investigated by polymerase chain reaction (PCR), polyacrylamide gel electrophoresis (PAGE) and denaturing high-performance liquid chromatography(DHPLC).

RESULTSTwo heteroduplexes and two homoduplexes in the PCR product from the heterozygous mutation of the exon 1 of KRT9 (497delAinsGGCT) were detected.

CONCLUSIONPCR products from KRT9 gene with heterozygous mutations contain two types of heteroduplexes. It is without the need to perform heating and cooling PCR products obtained from heterozygous mutations in advance before the mutation screening steps such as denaturing gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE), conformation-sensitive gel electrophoresis (CSGE), DHPLC and heteroduplex analysis (HA), etc.

Base Pair Mismatch ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; Electrophoresis, Polyacrylamide Gel ; Heteroduplex Analysis ; Heterozygote ; Humans ; Keratin-9 ; Keratins ; genetics ; Mutation ; Nucleic Acid Heteroduplexes ; Polymerase Chain Reaction

OBJECTIVETo analyze potential mutation of keration 9 gene (KRT9) in a Chinese family affected with epidermolytic palmoplantar keratoderma (EPPK) and to correlate genotype with the phenotype.

METHODSGenomic DNA was extracted from peripheral blood samples of 12 patients and 13 healthy individuals from the family and 100 unrelated individuals. Polymerase chain reaction (PCR) was used to amplify exons 1 and 6 of KRT9 gene. PCR products were sequenced bidirectionally in order to identify potential mutations.

RESULTSA heterozygous transversional mutation, 488G→A, was identified in exon 1 of KRT9 gene in all patients, which has resulted in substitution of a glutamine residue for arginine acid at position 163 (R163Q) of the KRT9 protein. The same mutation was not found in the 13 healthy members from the family and 100 unrelated individuals.

CONCLUSIONThe 488G→A mutation of KRT9 gene is probably the cause of EPPK in this Chinese family.

Adult ; Base Sequence ; DNA Mutational Analysis ; methods ; Female ; Humans ; Keratin-9 ; genetics ; Keratoderma, Palmoplantar, Epidermolytic ; genetics ; Male ; Molecular Sequence Data ; Mutation

OBJECTIVETo analyze the mutation of the keratin 9 gene (KRT9) in a pedigree with epidermolytic plamoplantar keratoderma (EPPK).

METHODSBlood samples were obtained from 4 affected and 3 normal individuals in this family. Mutation screening was carried out by polymerase chain reaction (PCR) and direct DNA sequencing.

RESULTSA heterozygous nucleotide C to T transition at position 484 in exon 1 of the KRT9 gene was detected in the 3 affected in this family, but was not found in normal individuals in the family and 100 unrelated individuals.

CONCLUSIONA missense mutation (484 C to T) in the KRT9 gene has been detected in this EPPK family, which is probably one of the molecular bases of the pathogenesis of the disease.

Adult ; Child, Preschool ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Humans ; Keratin-9 ; genetics ; Keratoderma, Palmoplantar ; genetics ; Male ; Molecular Diagnostic Techniques ; Mutation ; Mutation, Missense ; Pedigree

Adenocarcinoma ; metabolism ; pathology ; Adenoma ; pathology ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; CA-19-9 Antigen ; metabolism ; Cadherins ; metabolism ; Caroli Disease ; pathology ; Cholangiocarcinoma ; pathology ; Cystadenocarcinoma ; metabolism ; pathology ; Cystadenoma ; metabolism ; pathology ; Cysts ; pathology ; Diagnosis, Differential ; Hamartoma ; pathology ; Humans ; Keratin-19 ; metabolism ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Liver Diseases ; pathology

BACKGROUND: Epidermolytic palmoplantar keratoderma (EPPK) is an autosomal dominant disease of cornification which presents as severe thickening of the palms and soles with prominent epidermolytic hyperkeratosis pathologically. Recent studies have shown that EPPK is caused by mutations in the keratin 9 (K9) gene which is expressed essentially only in the palms and soles. Previously, We have reported that patients in one large pedigree of EPPK have an R162W substitution in the K9 protein. In this pedigree, two women whose husbands are both EPPK patients had become pregnant. OBJECTIVE: Since both women were concerned about this genetic disorder, we have performed prenatal diagnosis by biopsy analysis of chorionic villi tissue. METHODS: Chorionic villi biopsies were performed at 12 weeks gestation. Since the skin lesions are strictly confined to the palms and soles of the babies, the prenatal diagnosis of EPPK by ultrastructural analysis of fetal skin biopsy or amniotic fluid cells is highly problematic. Polymerase chain reaction amplification of specific allele (PASA) assay and direct DNA sequencing analyses were performed whether the fetuses carried mutant allele of K9 gene. RESULTS: PASA assay and direct DNA sequencing analyses showed that one fetus was normal, but the other fetus carried the abnormal allele. Subsequently, the mother of the unaffected fetus delivered a normal child, but the mother of the affected fetus terminated the pregnancy. CONCLUSION: We describe the analysis of the K9 mutation in the two fetuses at risk for EPPK. We believe that this is the first report of prenatal diagnosis for EPPK. But, we have to think about the ethical problems before we decide to perform the prenatal diagnosis of any kind of skin diseases.
Alleles ; Amniotic Fluid ; Biopsy ; Child ; Chorionic Villi ; Chorionic Villi Sampling ; Female ; Fetus ; Humans ; Hyperkeratosis, Epidermolytic ; Keratin-9 ; Keratoderma, Palmoplantar, Epidermolytic* ; Mothers ; Pedigree* ; Polymerase Chain Reaction ; Pregnancy* ; Prenatal Diagnosis* ; Sequence Analysis, DNA ; Skin ; Skin Diseases ; Spouses