Biomedical subjects

X Lee

Publications and source records attributed to X Lee.

27 records · Page 2Linked to original sources

PubMed1994

A beta 2RARE-LacZ transgene identifies retinoic acid-mediated transcriptional activation in distinct cutaneous sites.

Retinoic acid and its derivatives (retinoids) exert profound influences on epithelial growth differentiation in a variety of tissues, including the skin. How retinoic acid mediates these effects is not fully understood. The recent cloning of a series of nuclear receptors for retinoic acid (RARs) has demonstrated that these proteins can function as ligand-inducible transcriptional enhancing factors. Moreover, all receptors are members of the steroid/thyroid hormone multigene family. In vitro studies have demonstrated the expression of RAR alpha, RAR beta, and RAR gamma in various cell types found in the skin. While multiple isoforms exist for each of the three RARs, it is unclear where each of these receptors functions in vivo to mediate the tissue-specific effects of retinoic acid. As a first step in determining sites of retinoic acid-mediated transcriptional activation in the skin and its appendages, we developed a transgenic model in which the retinoic acid response element (RARE) of the RAR beta 2 isoform is linked to a beta-galactosidase reporter gene. Our observations consistently demonstrate that retinoic acid transcriptionally activates the beta 2RARE in distinct areas of the skin. Of interest, certain of these areas are known to contain stem cells. These data clearly demonstrate that this type of transgenic "reporter" model can be used to further define retinoic acid-regulated signal transduction pathways in the skin, as well as other complex tissues. Furthermore, these observations raise the possibility that transcriptional activation of RAR beta 2 may regulate the growth and differentiation programs of selected populations of stem cells in the skin and its appendages.

Animals↗

PubMed1994

Down-regulation of retinoic acid receptor beta in mammary carcinoma cell lines and its up-regulation in senescing normal mammary epithelial cells.

Retinoids are important cellular, dietary factors that regulate differentiation and cellular growth. They serve as ligands for specific nuclear receptors, the retinoic acid receptors (RARs). Ligand-activated receptors regulate gene transcription through target retinoic acid-responsive elements (RAREs) found in promoter regions. We have investigated the expression of retinoic acid receptor genes (alpha, beta, gamma) and retinoid X receptor beta in normal, senescing, and tumorigenic human mammary epithelial cells. We find that most tumor cells show a loss of RAR-beta expression, but that RAR-alpha and -gamma as well as retinoid X receptor beta are variably expressed in both normal and tumor cells. RAR-beta gene expression is induced both by retinoic acid and by fenretinide in normal cells, but tumor cells fail to respond to either. In contrast, RAR-beta expression increases with serial passage in senescing cells. Paradoxically, both normal and tumor cells can trans-activate an exogenous beta-RARE, as demonstrated by reporter gene assays. Oligonucleotide mobility shift assays with the beta-RARE show a single discrete complex in normal cells, whereas tumor cells exhibit a heterogeneous set of larger complexes, which indicates that tumor cells utilize a different array of factors within the beta-RARE. Reporter gene assays with extended promoter regions indicate the presence of negative regulatory elements and/or factor binding sites that reside between -1500 and the RARE located at -59, and that the promoter is down-regulated in MCF-7 tumor cells. Our findings reveal a dichotomy: RAR-beta transcription is down-regulated in tumor cells compared with normal human mammary epithelial cells, and up-regulated in senescence.

Base Sequence↗

PubMed1993

Crystallization and preliminary X-ray diffraction analysis of the complex of human placental 17 beta-hydroxysteroid dehydrogenase with NADP+.

Single crystals of human placental 17 beta-hydroxysteroid dehydrogenase, an enzyme that plays an important role in the interconversion of estrogens, were obtained as the NADP+ complex. These are the first crystals suitable for complete X-ray structure analysis ever reported for a steroid-converting enzyme from a human source. The crystals were grown by vapor diffusion at pH 7.5 with polyethyleneglycol (4000) as the precipitating agent. They have a monoclinic space group C2 and unit cell parameters are a = 123.03 A, b = 45.03 A, c = 61.29 A, and beta = 99.1 degrees. A complete set of diffraction data to 2.9 A has been collected on native crystals.

17-Hydroxysteroid Dehydrogenases↗

PubMed1993

Cultured human melanocytes express the intermediate filament vimentin.

Human melanocytes are neural crest-derived cells that synthesize the pigment melanin. These cells migrate from a central location to the dermal-epidermal junction early in gestation and situate themselves between keratinocytes of the basal layer of the epidermis, extending thin dendritic processes upwards into the epidermis. In vitro, neonatal melanocytes can assume a variety of morphologies, depending on the culture conditions. Using standard immunofluorescent, immunoblotting, and Northern blotting techniques, we investigated the expression of intermediate filament proteins and demonstrate here that cultured human melanocytes express vimentin gene and protein under a variety of culture conditions. Vimentin is a 57-kD intermediate filament protein synthesized primarily by cells of mesenchymal origin. It is transcribed as a single-messenger RNA species of 2.0 kb and the human gene is located on chromosome 10. As a member of the intermediate filament family of proteins, we suggest that vimentin is an important component of the cytoskeleton of neonatal, human melanocytes.

Blotting, Northern↗

PubMed1993

Clonal origin of dermatofibrosarcoma protuberans.

Dermatofibrosarcoma protuberans (DFSP) is a malignant tumor originating in the dermis. Although it is known to be locally aggressive, it only rarely metastasizes and will recur unless completely excised. The exact cell responsible for the development of a DFSP has been a matter of controversy for several decades; however, most histochemical and electron microscopic studies support a fibroblastic origin, with the tumor cells staining uniformly for vimentin and containing active endoplasmic reticulum synthesizing collagen. Cytogenetic analysis of some of these tumors has demonstrated at least two specific chromosomal abnormalities in DFSP and suggested that this tumor may be polyclonal in origin. To further address the clonal origin of this locally invasive, mesenchymal tumor, we analyzed DNA from two female patients by restriction fragment length polymorphisms and methylation analysis. Our data strongly support the concept that DFSP is monoclonal in origin and that this tumor mass reflects the clonal expansion of a single cell.

Adult↗

PubMed1993

DNA adducts as biomarkers for assessing exposure to polycyclic aromatic hydrocarbons in tissues from Xuan Wei women with high exposure to coal combustion emissions and high lung cancer mortality.

The high lung cancer rate in Xuan Wei, China, is associated with smoky coal use in unvented homes, but not with wood or smokeless coal use. Smoky coal combustion emits higher polycyclic aromatic hydrocarbon (PAH) concentrations than wood combustion. This study used DNA adducts as biomarkers for human exposure to PAH from combustion emissions. DNA adducts were determined by enzyme-linked immunosorbent assays (ELISA) in placentas and peripheral and cord white blood cells (WBC) from Xuan Wei women burning smoky coal or wood and from Beijing women using natural gas. Color ELISA gave positive results in 58, 47, and 5% of the placentas from Xuan Wei women burning smoky coal without and with chimneys, and from Beijing women, respectively. Fluorescence ELISA indicated that 46, 65, 56, and 25% of placentas were positive from Xuan Wei women who lived in houses without and with chimneys, Xuan Wei women burning wood, and Beijing controls, respectively. Peripheral WBC samples were positive in 7/9, 8/9, and 3/9 for the Xuan Wei women who lived in houses without and with chimneys and Beijing women, respectively. PAH-DNA adducts were detected in a higher percentage of placentas from Xuan Wei women living in houses exposed to smoky coal or wood emissions than from those of the Beijing controls. No dose-response relationship was observed between the air benzo[alpha]pyrene concentrations and DNA adduct levels or percentage of detectable samples. The results suggest that DNA adducts can be used as a qualitative biomarker to assess human exposure to combustion emissions.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

PubMed1990

Crystallization of recombinant rat cathepsin B.

A glycosylation-minus mutant of rat cathepsin B expressed in yeast has been purified and crystallized. X-ray diffraction data have been collected and molecular replacement for solving the structure is in progress. The space group for the recombinant rat cathepsin B was determined to be P2(1) with unit cell dimensions alpha = 62.2 A, b = 90.19 A, c = 47.07 A, and beta = 97.43 degrees. A unit cell contains 4 molecules and 2 molecules per asymmetric unit.

Animals↗

PubMed1989

Crystallization and preliminary X-ray diffraction studies of the complex of Maclura pomifera agglutinin with the disaccharide Gal beta 1-3GalNAc.

Single crystals of Maclura pomifera agglutinin, a seed lectin from the Moraceae family, complexed with the disaccharide Gal beta 1-3GalNAc have been obtained by the method of vapor diffusion with Li2SO4 as precipitant at pH 4.5. The crystals belong to the trigonal space group P3(1)21 or P3(2)21, with a = b = 67.4 A, c = 149.3 A. They contain two subunits per asymmetric unit and diffract beyond 2.7 A. This and other evidence indicate that both this lectin and the Artocarpus integrifolia lectin, jacalin, have dimeric structures rather than the tetrameric structures previously proposed.

Antigens, Surface↗

PubMedDate not supplied

Identification of inosine as an endogenous modulator for the benzodiazepine binding site of the GABAA receptors.

Previously we have reported the presence of endogenous ligands that are involved in the regulation of the binding of muscimol to the GABA binding site of the GABAA receptors. Here, we report the presence of multiple forms of endogenous ligands in the brain which modulate the binding of flunitrazepam (FNZP) to the benzodiazepine (BZ) binding site of the GABAA receptor. Furthermore, one of the endogenous ligands for the BZ receptors, referred to as EBZ, has been identified as inosine based on the following observations: (1) standard inosine and the EBZ have identical NMR and UV spectra; (2) the elution profile of inosine and the EBZ from a HPLC column are indistinguishable, and (3) inosine and the EBZ show identical activity in inhibiting [3H]FNZP binding.

Animals↗