PURPOSE: Dysplasia or flat adenoma of the stomach is regarded as a precancerous lesion. However, the frequency and the evolutionary process of malignant transformation of gastric dysplasia are still debated. In order to see whether the lesion was a monoclonal or a polyclonal proliferation, clonality was assayed by X-linked HUMARA polymorphism. MATENRIALS AND METHODS: DNA was extracted from the paraffin-embedded tissue of 16 consecutive cases of endoscopic biopsy, eight of which supplied both dysplastic and nondysplastic tissue for comparison. HUMARA was amplified by PCR with or without pretreatment with methylation- sensitive restriction enzyme, HpaII. The amplification products were electrophoresed on polyacrylamide gel and silver-stained. RESULTS: Among the 16 cases, 13 cases were informative and 3 cases noninformative. Of the 13 cases, one case showed skewed lyonization, rendering 12 cases to be analyzed further. A monoclonal band pattern was noted in 2 cases, and a polyclonal band pattern in 10 cases. A review of the histopathologies of the monoclonal and the polyclonal cases did not reveal features discriminating the two groups. CONCLUSION: These results suggest that gastric dysplasia is a disease entity heterogeneous in the genetic level, and many cases may be non-neoplastic.
Adenoma ; Biopsy ; DNA ; Polymerase Chain Reaction ; Stomach* ; X Chromosome Inactivation

Hemophilia is a relatively rare bleeding disorder. It is an X-linked hereditary bleeding disorder caused by a deficient or defective coagulation factor VIII (Hemophilia A) or factor IX (Hemophilia B). Hemophilia A is more common than Hemophilia B. The X-linked inheritance pattern results in men expressing the disease and women typically being carriers. Under rare circumstances a woman can also show a bleeding phenotype.

A 13 year-old female presented with profuse vaginal bleeding. She had history of several hospital admissions because of bleeding manifestations like hematuria and epistaxis. Based on the pedigree analysis and results of factor IX assay tests she was diagnosed to have Hemophilia B of moderate severity. She was given hormonal and non-hormonal treatments as well as blood transfusions which stop the bleeding and corrected the anemia. A multidisciplinary approach of management involving the gynecologist, hematologist and a geneticist will be beneficial to the patient.

The inheritance, clinical manifestations, diagnosis and treatment of Hemophilia B in a female adolescent are discussed

Human ; Female ; Adolescent ; Hemophilia B ; X Chromosome Inactivation ; Uterine Hemorrhage

OBJECTIVETo explore the pathogenetic mechanism for a female patient affected with hemophilia A (HA).

METHODSPotential genetic defect was detected with inverse shifting-polymerase chain reaction (IS-PCR). The pattern of X chromosome inactivation was determined with a human androgen receptor assay (HUMARA assay). G-banded karyotyping was carried out to exclude potential chromosome aberrations.

RESULTSIS-PCR showed that the defect of FVIII gene was the distal type of intron 22 inversion. The HUMARA assay showed that the X chromosome inactivation was non-random, and that the mother's X chromosome activity was lower than that of the father's X chromosome which has carried the inverted FVIII gene. No abnormalities were found with G-banded chromosomes.

CONCLUSIONThe prevalence of female HA patient may be caused by non-random inactivation of X chromosomes.

Adolescent ; Female ; Hemophilia A ; etiology ; genetics ; Humans ; Karyotyping ; Polymerase Chain Reaction ; Receptors, Androgen ; analysis ; X Chromosome Inactivation

Long non-coding RNAs (lncRNAs) reportedly function as important modulators of gene regulation and malignant processes in the development of human cancers. The lncRNA JPX is a novel molecular switch for X chromosome inactivation and differentially expressed JPX has exhibited certain clinical correlations in several cancers. Notably, JPX participates in cancer growth, metastasis, and chemoresistance, by acting as a competing endogenous RNA for microRNA, interacting with proteins, and regulating some specific signaling pathways. Moreover, JPX may serve as a potential biomarker and therapeutic target for the diagnosis, prognosis, and treatment of cancer. The present article summarizes our current understanding of the structure, expression, and function of JPX in malignant cancer processes and discusses its molecular mechanisms and potential applications in cancer biology and medicine.
Humans ; RNA, Long Noncoding/genetics* ; Neoplasms/genetics* ; MicroRNAs/genetics* ; Gene Expression Regulation ; X Chromosome Inactivation

To investigate the XIST gene expression and its effect in a Klinefelter''s patient, we used Klinefelter''s syndrome (XXY) patient with azoospermia and also used a normal male (XY) and a normal female (XX) as the control, We were performed cytogenetic analysis, Y chromosomal microdeletion assay (Yq), semi-quantitative RT-PCR, and the Northern blot for Klinefelter''s syndrome (KS) patient, a female and a male control, We extracted total RNA from the KS patient, and from the normal cells of the female and male control subjects using the RNA prep kit (Qiagen), cDNA microarray contained 218 human X chromosome-specific genes was fabricated. Each total RNA was reverse transcribed to the first strand cDNA and was labeled with Cy-3 and Cy-5 fluorescein, The microarray was scanned by ScanArray 4000XL system. XIST transcripts were detected from the Klinefelters patient and the female by RT-PCR and Northern blot analysis, but not from the normal male, In the cDNA microarray experiment, we found 24 genes and 14 genes are highly expressed in KS more than the normal male and females, respectively. We concluded that highly expressed genes in KS may be a resulted of the abnormal X inactivation mechanism.
Azoospermia ; Blotting, Northern ; Cytogenetic Analysis ; DNA, Complementary* ; Female ; Fluorescein ; Gene Expression ; Humans ; Klinefelter Syndrome* ; Male ; Oligonucleotide Array Sequence Analysis* ; RNA ; X Chromosome Inactivation* ; X Chromosome*

OBJECTIVERett syndrome (RTT) is a neurodevelopmental disorder occurring almost exclusively in females as sporadic cases due to de novo mutations in the methyl-CpG-binding protein 2 gene (MECP2). Familial cases of RTT are rare and are due to X-chromosomal inheritance from a carrier mother. Recently, DNA mutations in the MECP2 have been detected in approximately 84.7% of patients with RTT in China. To explain the sex-limited expression of RTT, it has been suggested that de novo X-linked mutations occur exclusively in male germ cells resulting therefore only in affected daughters. To test this hypothesis, we have analyzed the parental origin of mutations and the XCI status in 15 sporadic cases with RTT due to MECP2 molecular defects.

METHODSAllele-specific PCR was performed to amplify a fragment including the position of the mutation. The allele-specific PCR products were sequenced to determine which haplotype contained the mutation. It was then possible to determine the parent of origin by genotyping the single nucleotide polymorphism (SNP) in the parents. The degree of XCI and its direction relative to the X chromosome parent of origin were measured in DNA prepared from peripheral blood leucocytes by analyzing CAG repeat polymorphism in the androgen receptor gene (AR).

RESULTSExcept for 2 cases who had a frameshift mutation; all the remaining 13 cases had a C-->T transition mutation. Paternal origin has been determined in all cases with the C-->T transition mutation. For the two frameshift mutations, paternal origin has been determined in one case and maternal origin in the other. The frequency of male germ-line transmission in mutations is 93.3%. Except for 2 cases who were homozygotic at the AR locus, of the remaining 13 cases, 8 cases had a random XCI pattern; the other five cases had a skewed XCI pattern and they favor expression of the maternal origin allele.

CONCLUSIONDe novo mutations in sporadic RTT occur almost exclusively on the paternally derived X chromosome and that this is most probably the cause for the high female: male ratio observed in sporadic cases with RTT. Random XCI was the main XCI pattern in sporadic RTT patients. The priority inactive X chromosome was mainly of paternal origin.

Chromosome Aberrations ; Chromosomes, Human, X ; Female ; Humans ; Male ; Methyl-CpG-Binding Protein 2 ; genetics ; Mutation ; Polymorphism, Single Nucleotide ; Rett Syndrome ; genetics ; X Chromosome Inactivation

BACKGROUND: While neurofibromas have generally been regarded as polyclonal hyperplastic lesions, it remains unclear whether the tumor is a true neoplasm or a hyperplastic lesion. METHODS: Determination of clonality by X chromosome inactivation pattern was investigated in twenty-one cases of neurofibroma employing enzyme digestion and PCR of the HUMARA gene. The histological, immunohistochemical, and ultrastructural characteristics of the tumors were also examined. RESULTS: Immunohistochemically, most of the tumor cells showed vimentin and S-100 protein positivity. Axons were demonstrated by neurofilament protein positivity and were seen mainly at the periphery and rarely in the central portion of the tumor. Ultrastructurally, the tumors were composed of a variety of cell types: perineurial cells, Schwann cells, fibroblasts, and axons. X chromosome inactivation analysis was completed on thirteen out of fifteen cases in which DNA was successfully extracted. Of thirteen neurofibromas that were heterozygous at the HUMARA loci, eleven showed a polyclonal pattern. The remaining two cases were considered as indeterminate for clonality because of unequal band intensity and failure to obtain the normal control DNA. CONCLUSION: The results from this study suggest that neurofibromas are polyclonal in origin and might be a neoplastic lesion comprising non-neoplastic cells among constituent components.
Axons ; Digestion ; DNA ; Fibroblasts ; Immunohistochemistry ; Neurofibroma* ; Polymerase Chain Reaction* ; S100 Proteins ; Schwann Cells ; Vimentin ; X Chromosome Inactivation

OBJECTIVETo investigate the role of clonality analysis by X-chromosome inactivation in the diagnosis of cervical lymph node metastasis from squamous carcinoma of the head and neck.

METHODSTwenty cases of clinical NOM0 squamous carcinoma of the head and neck with either pathologically confirmed or suspected occult micrometastasis in the cervical lymph node were recruited. Interested DNA samples were procured through tissue microdissection and one-step proteinase K digestion, and the clonality analysis was carried out by means of restriction enzyme digestion and amplification of human androgen receptor markers (HUMURA) to check out the status of X-chromosome inactivation. The clonal origin of the primary tumor cells and the interested cell clones in the cervical lymph node was traced by X-chromosome inactivation, and the diagnosis of cervical lymph node micrometastasis was either confirmed or ruled out.

RESULTSTumor cells from both primary and metastatic lesions were monoclonal and identical in clonal origin in 10 patients with pathologically confirmed cervical lymph node metastasis, whose metastatic tumor cells expressed EGF receptor. For 10 patients with suspected micrometastasis in the neck nodes, whose focused lesions did not expressed any EGF receptor protein by immunohistochemistry, the identical and monoclonal origin between the primary tumor and the suspected metastatic lesion in the neck node was confirmed in 6 patients, and the polyclonal origin of the neck node lesions was revealed in other 4 patients. The diagnosis of micrometastasis in the neck node was thus ascertained in 6 and ruled out in 4 suspected cases.

CONCLUSIONSExamination of X-chromosome inactivation pattern is a useful method for identification of the neck node occult micrometastasis from squamous carcinoma of the head and neck.

Carcinoma, Squamous Cell ; diagnosis ; pathology ; Female ; Head and Neck Neoplasms ; diagnosis ; pathology ; Humans ; Lymphatic Metastasis ; diagnosis ; Middle Aged ; X Chromosome Inactivation

OBJECTIVE: The XIST gene is considered to be an attractive candidate gene for skewed X-chromosome inactivation and a possible cause of primary ovarian insufficiency (POI). The purpose of this study was to investigate whether the XIST gene promoter mutation is associated with idiopathic POI in a sample of the Korean population. METHODS: Subjects consisted of 102 idiopathic POI patients and 113 healthy controls with normal menstrual cycles. Patients with the following known causes of POI were excluded in advance: cytogenetic abnormalities, prior chemo- or radiotherapy, or prior bilateral oophorectomy. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: The mean age of onset of ovarian insufficiency was 28.7+/-8.5 years and the mean values of serum luteinizing and follicle-stimulating hormones and estradiol in the POI group were 31.4+/-18.2 mIU/mL, 74.5+/-41.1 mIU/mL, and 30.5+/-36.7 pg/mL, respectively. We found no cytosine to guanine (C43G) variation in the XIST gene in both POI patients and controls. CONCLUSION: The C43G mutation in the promoter region of the XIST gene was not present in the Korean patients with idiopathic POI in our study, in contrast to our expectation, suggesting that the role of XIST in the pathogenesis of POI is not yet clear.
Age of Onset ; Chromosome Aberrations ; Cytosine ; Estradiol ; Female ; Guanine ; Humans ; Lutein ; Menstrual Cycle ; Ovariectomy ; Primary Ovarian Insufficiency* ; Promoter Regions, Genetic* ; Radiotherapy ; X Chromosome Inactivation

A 31-year-old woman, who was pregnant with twins, underwent chorionic villus sampling because of increased nuchal translucency in one of the fetuses. Cytogenetic analysis showed a normal karyotype in the fetus with increased nuchal translucency. However, the other fetus, with normal nuchal translucency, had a derivative X chromosome (der(X)). For further analysis, fluorescence in situ hybridization (FISH) and additional molecular studies including fragile X analysis were performed. FISH analysis confirmed that the Y chromosome was the origin of extra segment of the der(X). The X-chromosome breakpoint was determined to be at Xq27 by FMR1 CGG repeat analysis, and the Y-chromosome breakpoint was determined to be at Yq11.23 by the Y chromosome microdeletion study. To predict the fetal outcome, the X-inactivation pattern was examined, and it revealed non-random X inactivation of the der(X). To the best of our knowledge, the identification of an unbalanced Xq;Yq translocation at prenatal diagnosis has never been reported. This study was performed to identify precise breakpoints and the X-inactivation pattern as well as to provide the parents with appropriate genetic counseling.
Adult ; Chorionic Villi Sampling ; Cytogenetic Analysis ; Female ; Fetus* ; Fluorescence ; Genetic Counseling ; Humans ; In Situ Hybridization ; Karyotype ; Nuchal Translucency Measurement ; Parents ; Pregnancy ; Prenatal Diagnosis ; Twins ; X Chromosome ; X Chromosome Inactivation ; Y Chromosome