ABSTRACT
Cornified envelope is highly insoluble structure formed beneath the plasma membrane during terminal differentiation of keratinocytes and is stabilized by cross linking of various proteins, including involucrin, loricrin, and cornifin. Psoriasis is a chronic skin disease characterizing inflammatory reaction and hyperproliferation of keratinocyte. There are some differences in involucrin immunolabelling in stratum corneum between normal and psoriasis epidermis. Labelling was convergent to cornified envelope in psoriasis skin but throughout cytoplasm in normal skin. To compare terminal differentiation patterns of normal and psoriasis keratinocytes, we reconstructed normal and psoriatic artificial skin by using primary cultured keratinocytes from normal and psoriasis skin and then performed immunogold labelling for involucrin in stratum corneum. Psoriatic artificial skin had thin and poorly organized corneal layer. Immunogold labelling for involucrin revealed same pattern of that in vivo by showing throughout cytoplasm in lower layer but convergent cornified envelope in upper layer. Compared with psoriatic artificial skin, normal artificial skin had well organized and thick stratum corneum. Involucrin labelling was throughout cytoplasm in most of corneal layer but convergent to cornified envelope in some uppermost cells. Even though some cells show convergent pattern in normal artificial skin, absolute number of this pattern was no lesser than in artificial psoriatic skin because of normal artificial skin had thick stratum corneum. This result showed there was no difference in involucrin distribution in terminal differentiation of normal and psoriasis keratinocytes in organotypic culture model. It is concluded that although well organized multiple corneal layers are formed in normal artificial skin, they can not reach to full maturation of cornified envelope, and difference of involucrin localization in cornified envelope of psoriasis epidermis is related with not peculiarities of the cells but rapid growing in vivo.
Subject(s)
Cell Membrane , Cytoplasm , Epidermis , Keratinocytes , Psoriasis , Skin , Skin Diseases , Skin, ArtificialABSTRACT
BACKGROUND: Insulin-dependent diabetes mellitus (IDDM) is caused by the autoimmune destruction of pancreatic beta-cells. Susceptibility to IDDM appears to depend on more than one genetic locus. Evidence of a genetic linkage for IDDM2 was found in male meioses from French and North American populations. It is linked to maternal imprinting (i.e. monoalleleic expression of the insulin gene) that is considered the most likely cause of these gender-related differences. IGF2 is expressed only in the paternal allele and, therefore, is considered a candidate gene for IDDM2 transmission because of its important autocrine/paracrine effects on the thymus, lymphocytes and pancreas. Nevertheless, it remains controversial whether the parental origin of IDDM2 influences IDDM susceptibility. METHODS: Using PCR and semi-quantitative RT-PCR, we analyzed the INS/ PstI+1127 and IGF2/ApaI polymorphisms and RNA expression level between PstI (+/-) and PstI (+/+) to determine genotype and allele-specific expression of the INS and IGF2 genes. RESULTS: INS/PstI (+/+) and IGF2/ApaI (+/-) were observed in 36 (97.3%) of 37 IDDM patients and in 29 (72.5%) of 40 IDDM patients, respectively. The presence of both IGF2 alleles in RNA was observed in 21 (91.6%) of 24 IDDM patients. Our results show a 3-fold increase in RNA expression from PstI (+/-) allele over PstI (+/+) allele. CONCLUSION: Our conclusion does not entirely exclude IGF2 as the gene involved in IDDM2, even though the parental effect of IDDM2 transmission is not related to IGF2 maternal imprinting. The INS genotype appeared mostly in the PstI (+/+) homozygote and, therefore, we could not explain the INS imprinting pattern in Korean type 1 diabetic patients. Genetic differences between populations may account for the discrepancy between Korean type I diabetic patients and American or French type I diabetic patients.
Subject(s)
Adolescent , Child , Female , Humans , Male , Diabetes Mellitus, Type 1/genetics , Genomic Imprinting , Insulin/genetics , Insulin-Like Growth Factor II/genetics , Korea , Sex FactorsABSTRACT
Human uniparental gestations such as gynogenetic ovarian teratomas provide a model to evaluate the integrity of parent-specific gene expression - i.e. imprinting - in the absence of a complementary parental genetic contribution. The few imprinted genes characterized so far include the insulin-like growth factor-2 gene (IGF2) coding for a fetal growth factor and H19 gene whose normal function is unknown but it is likely to act as an mRNA. IGF2 is expressed by the paternal allele and H19 by the maternal allele. This reciprocal expression is quite interesting because both H19 and IGF2 genes are located close to each other on chromosome 11p15.5. In situ RNA hybridization analysis has shown variable expression of the H19 and IGF2 alleles according to the tissue origin in 11 teratomas. Especially, Skin, derivative of ectoderm, is expressed conspicuously. We examined imprinting of H19 and IGF2 in teratomas using PCR and RT-PCR of exonic polymorphism. H19 and IGF2 transcript could be expressed either biallelically or monoallelically in the teratomas. Biallelic expression (i.e., loss of imprinting) of IGF2 occured in 5 out of 6 mature teratomas and 1 out of 1 immature teratoma. Biallelic expression of H19 occured in 4 out of 10 mature teratomas and 1 out of 1 immature teratoma. Expression levels of H19 and IGF2 transcript using the semi-quantitative RT-PCR had no relation between monoallelic and biallelic expression. Moreover, IGF2 biallelic expression did not affect allele-specificity or levels of H19 expression. These results demonstrate that both genes, H19 and IGF2, can be imprinted, expressed and regulated independently and individually of each other in ovarian teratoma.
Subject(s)
Humans , Alleles , Clinical Coding , Ectoderm , Exons , Fetal Development , Gene Expression , Genomic Imprinting , Insulin-Like Growth Factor II , Parents , Polymerase Chain Reaction , RNA , RNA, Messenger , Skin , TeratomaABSTRACT
To investigate fragile sites induced by aphidicolin which is a specific inhibitor of eukaryotic DNA polymerase a which is primarily associated with chromosomal DNA replication in human lymphocytes, HaCat cells (human keratinocytes) and MRC-5 cells (human embryonic lung fibroblast), we cultured each cells in RPMI 1640 with 10% fetal calf serum and 2% PHA. Treatment of the cells with aphidicolin was generally carried out for the last 24 hours of culturing. The drug was dissolved in DMSO and used at final concentrations of 0.05~0.15 mg/ml, corresponding to a maximum DMSO concentration of 0.028%. Karyotypes of each cells were performed by routine method, and 50 metaphases were scored for each culture for analysis of breakage rate. Experimental cells treated with APC showed a dose dependent sensitivity and the amounts of chromosome breakage induced by APC are the highest in concentration of 0.15 mg/ml. The frequency of fragile sites on each cells appeared in MRC-5 cells, lymphocytes and HaCat cells in order. The common fragile sites on all experiments was 16q23, and the common fragile sites on embryonic cells was 1p31. It can be concluded that gene or nucleic acid which is located on 16q23 is the most important factor to induce chromosomal breakage with sensitivity to aphidicolin and 1p31 is important site to induce chromosomal breakage in embryonal cells.
Subject(s)
Humans , Aphidicolin , Chromosome Breakage , Dimethyl Sulfoxide , DNA , DNA Replication , Karyotype , Lung , Lymphocytes , MetaphaseABSTRACT
Human uniparental gestations such as androgenetic hydatidiform moles provide a model to evaluate the integrity of parent-specific gene expression,-i.e, genomic imprinting,- in the absence of a complementary parental genetic contribution. Several imprinted genes are characterized so far including the insulin-like growth factor-2 gene (IGF2) coding for a fetal growth factor and the Hl9 gene whose normal function is unknown but which is likely to act as an untranslated mRNA for its tumor-suppressing function. IGF2 is expressed exclusively from the paternal allele while Hl9 from the maternal allele. Such an alternate expression is quite interesting because both Hl9 and IGF2 genes are located close to each other on chromosome 11p15.5. An in situ hybridization analysis has shown strong expression of Hl9 and IGF2 alleles in nine hydatidiform moles. Especially, a prominent expression of Hl9 and IGF2 was detected in cytotrophoblast and the cellular localization was almost paralleled in Hl9 and IGF2 transcripts . Hl9 and IGF2 genes could be expressed either biallelically or monoallelically in the moles. However, IGF2 biallelic expression did not affect allele-specificity of Hl9 expression.. These results suggest that both H19 and IGF2 transcripts are expressed in the same cells and that the functional imprinting of H19 and IGF2 genes in hydatidiform moles can be controlled individually and independently of each other.
Subject(s)
Female , Humans , Pregnancy , Alleles , Chromosome Aberrations , Clinical Coding , Fetal Development , Genomic Imprinting , Hydatidiform Mole , In Situ Hybridization , Insulin-Like Growth Factor II , Parents , RNA, Messenger , TrophoblastsABSTRACT
Since Steeg, et al.(1988) identified NM23/NDP kinase as non -metastasis gene, other multiple functions of have reported. One of them, Postel, et al.(1993) suggested that transcription factor PuF, being encoded by NM23 -H2/NDP kinase gene, interacts with nuclease hypersensitive element located upstream of the c -myc gene. C -myc amplification and activation can be present in squamous cell carcinoma of the head and neck as well as in an increased metastatic propensity for individual tumor. To clarify the role of NM23/NDP kinase on c -myc expression, comparison of these two gene expressions in cell lines was done. No direct correlation of expression kinetics was found. A plasmid containing human c -myc fragment was cloned upstream of chloramphenicol acetyltransferase (CAT) gene. When murine melanoma cell line was cotransfected with a murine NM23 -M2 including expression vector and c -myc CAT, CAT activity was elevated, while no change of CAT activity was found in the cotransfectant of human NM23 -H2 and c -myc CAT. Data suggest that murine NM23 -M2 gene transactivates c -myc gene indirectly with a cellular factor in murine cell line which dose not work with human NM23 -H2 gene. Additionally, we found same kinetics of NM23 -H2/NDP kinase and c -myc expression change correlated with proliferation of PLC/PRF/5 which was induced by HGF.
Subject(s)
Animals , Cats , Humans , Carcinoma, Squamous Cell , Cell Line , Chloramphenicol O-Acetyltransferase , Clone Cells , Gene Expression , Head , Kinetics , Melanoma , Neck , Phosphotransferases , Plasmids , Transcription Factors , Transcriptional ActivationABSTRACT
PURPOSE: Human herpesviruses have been associated with the etiology of several human cancers. The role of these viruses in carcinogenesis has not yet been clarified. This study focused on identifying and characterizing the transforming potential of cloned DNA fragments from human cytomegalovirus (HCMV). MATERIALS AND METHODS: Multiple DNA fragments of HCMV were applied to cells for transformation. Morphological transforming region II (mtrII) of HCMV strain Towne has been identified to a 3.0kb XbaI-BamHI DNA fragment which was retained in transformed cells. The transforming activity was induced by a 980 bp BaII-Xho I subfragment (pBS980) containing both promoter/ regulatory elements as well as three open reading frames (ORFs), i.e., 79ORF, 83ORF, and 34ORF. The ORFs have been evaluated for transforming potential in NIH3T3 cells. RESULTS: MtrII (pBS980) has BglII restriction enzyme site which divides into two subfragments, pBS440 and pBS540, the latter has whole 83ORF, 34ORF, and fragment of 79ORF, the former has only fragment of 79ORF. Among three ORFs, 83ORF and 34ORF were not functional in transformation, because in pBS540 these ORFs were not truncated. CONCLUSION: The 79ORF (79-aa transforming peptide) has allowed a better approach to determine the role of HCMV in human carcinogenesis.
Subject(s)
Animals , Humans , Carcinogenesis , Clone Cells , Cytomegalovirus , DNA , Ecthyma, Contagious , Herpesviridae , Open Reading FramesABSTRACT
To understand the role of IGF2 gene in development of human ovary, IGF2 expression was detected by monoclonal antibody for IGF2 to its producted protein with immunohistochemical technique on human ovarian tissues. The results was as follows. IGF2 was highly expressed in ovum of mature follicle, IGF2 expression, however, was not high in granulosa and the cells. IGF2 was not highly expressed in ovum of primary follicle. Highly expressed IGF2 was found on corpus luteum and no expression of IGF2 was found in stroma and epithelial cells. These results suggest that IGF2 is important role in ovulation and in production of progesterone. Abnormal IGF2 expression may be concerned to carcinogenesis of ovarian tumor because most of all tumor from ovary is originated from epithelium.
Subject(s)
Female , Humans , Carcinogenesis , Corpus Luteum , Epithelial Cells , Epithelium , Ovary , Ovulation , Ovum , Progesterone , Theca CellsABSTRACT
Cytogenetic techniques were used to detect specific chromosomal losses and / or stuctural changes in 6 meningioma cell population of 11meningioma patients. Polymorphic DNA markers were uti.lized to investigate the loss of constitutional heterozygosity on chromosomes 8. 17 and 22 in 9 meningioma cell population of 1l meningioma patients. As a result, 5 cases(M-2.4,5.9, and 10) represented 45. XX. -22 or 45, XY.-22 as stem line. In addition to chromosome 22, other chromosomes were lost randomly. In one case(M-3) normal karyotypic pattern was oberved. The 9q+ structural change was also noted in case M-2. This structural change was thought to be the chromosomal involvement secondary to the loss of chromosome 22 in meningioma. Retentions of constitutional heterozygosity on chromosomes 8 and 17 were found in all cases. Loss of constitutional hererozygosity on chromosome 22 were found at Hind m RFLP of v-sis in cases M-1 and M-7. EcoRI RFLP of v-sis in case M-1. Bgl II RFLP of v-sis case M-1. Xba I RFLP of v-sis in cases M-6. M-9 and M-11. And EcoRI RFLP of bcr in all cases. Rearrangement of chromosome 22 in case M-1 was detected on the Xba I RFLP of v-sis as extra band(3.14kb). The reduction to hemizygosity on chromosome 22 was one important step in tumorigenesis of meningioma. Monosomy 22 might operate at the primary level of tumor initiation. Random losses of other chromosomes or structural changes as 9q+ were postula!ed to be related to tumor development.
Subject(s)
Humans , Carcinogenesis , Chromosomes, Human, Pair 22 , Cytogenetic Analysis , Cytogenetics , Genetic Markers , Meningioma , Monosomy , Polymorphism, Restriction Fragment LengthABSTRACT
No abstract available.
Subject(s)
Cytogenetic Analysis , Cytogenetics , Urinary Bladder Neoplasms , Urinary BladderABSTRACT
No abstract available.
ABSTRACT
No abstract available.
Subject(s)
Animals , Cricetinae , Humans , Ammonium Compounds , Ammonium Sulfate , Bromodeoxyuridine , CHO Cells , Chromosome Aberrations , Siblings , Sister Chromatid ExchangeABSTRACT
The XX-male or sex reversal syndrome is a rare entity, which a is phenotypic man with a 46, XX female karyotype. Since it was first reported by la Chapelle and associates in 1964, more than 150 XX males have been reported. Recently we experienced a 18-year-old XX-male with gynecomastia and hypospadias. Clinical, endocrinological and genetically studies were presented and theories regarding the etiology of the XX-male syndrome were discussed with review of literatures.